| 00:00 | I Ok folks, welcome um so I assume you got my email |
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| 00:37 | back in the routine again? Blackboard this week. Smart work next |
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| 00:44 | So stuff you on monday of next , so be back in that mode |
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| 00:49 | a bit. The next exam is for a while because we got spring |
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| 00:54 | um Kind of uh in the middle . So basically March 24 25. |
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| 01:03 | a long ways away. We'll be Unit three by then. So uh |
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| 01:12 | , so you started going to finish viruses part one and then get into |
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| 01:17 | two, which is mainly focusing on viral life cycles. Okay, um |
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| 01:30 | 13 is one of those class but it's not gonna get much into |
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| 01:37 | . We do a little bit, not a lot. Most of that |
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| 01:40 | be left for the following Tuesday, is uh where we at right |
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| 01:46 | so we'll do some here and then it up um uh in the next |
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| 01:52 | here. Okay. Um Okay, regarding the exam overall, I |
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| 02:00 | I was pleased with it. I this yesterday on blackboards, so the |
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| 02:07 | , so I do like seeing lots fat bars on the right side. |
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| 02:13 | so that's my goal, always my . So, but you know, |
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| 02:19 | 70 rounding up 71 average, but know, that's pretty much what I |
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| 02:27 | to hit is somewhere 70 plus plus minus two points. Give or |
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| 02:33 | Okay, so um I am, what I'm about to say is going |
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| 02:43 | be applicable to probably focus over on side. Okay, hold on this |
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| 02:50 | over here, over here. so number one um you don't don't |
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| 03:00 | crazy. Okay, calm down. right too. Is um you will |
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| 03:07 | be asked any more questions on unit material because the final exam is not |
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| 03:15 | . Just each exam is its own covering that previous quarter of the |
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| 03:20 | Okay, so that's number two. three is whatever you did that got |
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| 03:26 | to this. Not so great Do not repeat that process again. |
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| 03:31 | . If you do, I will you guaranteed. I'll put my hope |
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| 03:38 | my money on you will make the grade next time around, guaranteed. |
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| 03:45 | . Um And so don't do that . People will think, okay, |
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| 03:50 | made a not so good grade. me study twice as hard now. |
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| 03:56 | , I'm gonna study twice as hard the same blank blank study method. |
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| 04:02 | again, guaranteed disaster. Okay, you've got to change them. All |
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| 04:07 | , so um I will I'm not get the whole steel now, but |
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| 04:14 | will just snip it on day one 1 17 23 that I will send |
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| 04:21 | link on the email this week on to study at least at least my |
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| 04:27 | on how to do it. There was a person. Is he |
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| 04:29 | today? Is he at the guy he came by, I don't want |
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| 04:35 | name. I don't know your You left your headphones in the lab |
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| 04:40 | open the door there are he followed . He did very well. |
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| 04:47 | okay. So you have to listen me. Here's one of your |
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| 04:52 | Okay. So anyway, but that of the lecture, right? That's |
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| 04:57 | you follow. Okay. Perfect. . So, um anyway, so |
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| 05:02 | at that. You got questions obviously me know. Okay, so uh |
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| 05:08 | you want to look at the just face to face virtual and we |
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| 05:15 | go over it. Okay, So , so I'll leave it at |
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| 05:19 | So, the other thing, the important thing is uh I think even |
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| 05:24 | no matter how you perform have amnesia amnesia because now your focus is um |
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| 05:34 | right here is this now your is that okay? So forget everything |
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| 05:40 | . Now that so um having said not gonna ask specific questions about, |
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| 05:47 | know, more material, you know some of that those concepts we're gonna |
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| 05:54 | over into 13 and 14 metabolism. talked about growth and And Blah |
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| 06:02 | Right. That's all gonna be applicable uh 13 or 14. Okay. |
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| 06:10 | um any questions? All right, . Okay. Okay. All |
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| 06:22 | So let's I think we just have little bit of a recap. Um |
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| 06:27 | obviously we're talking about viruses. Like viruses um Part one was about |
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| 06:34 | defining a virus structure. Just some the basic features basic life cycle. |
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| 06:40 | . So he was just showing you of summarizing a definition of a you |
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| 06:46 | , they're not selves they are uh we would have right. They obviously |
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| 06:56 | uh structurally size side branches then um capsule rights that they have at minimum |
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| 07:04 | viruses have this protein covering that covers genome uh shapes and be symmetrical |
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| 07:16 | This kind of more kind of this form tailed viruses um making the envelopes |
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| 07:25 | refers to whether it's required an envelope the host cell um as it exits |
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| 07:31 | captain exits that kind of wraps around it can be without without a |
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| 07:36 | Okay. And then um so viral proteins in the search from infectivity recognition |
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| 07:46 | some of those others have different They often look like very prominent |
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| 07:53 | So um yeah, but the point is you're gonna they're going viral specific |
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| 08:00 | on the surface. Okay. Uh or whatever. All right. And |
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| 08:07 | we looked at the reproduction rights. the basics. And so we're getting |
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| 08:13 | more specifics uh viral types differ in process but pretty much overall this is |
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| 08:22 | . And we know that there's gonna variation. Right? So from the |
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| 08:25 | , of course they get resources depending the viral type. They may or |
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| 08:30 | not need to use the host but delivered opponents. They will need uh |
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| 08:38 | . So I one of them as get into. Not not so much |
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| 08:43 | but likely thursday is RNA viruses that kind of the ones that typically uh |
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| 08:49 | look problematic helps to kind of uh that. Okay, this is what's |
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| 08:56 | in, right? And ultimately this what's coming out. Right? So |
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| 09:00 | heart on the part of the it's going to make it a |
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| 09:04 | It's gonna be making ultimately making viral that exits the cell. Right? |
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| 09:10 | what's going on in between is everything allow that to happen? And what |
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| 09:15 | you need to look at the virus ? Right. It needs proteins, |
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| 09:20 | proteins, right, captured proteins and else that's there. And so I |
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| 09:25 | to assemble this because what's happening is just having a genome in here, |
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| 09:32 | ? You have to put everything else it to make viral particles. So |
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| 09:35 | have to copy that you have to proteins, you gotta assemble, put |
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| 09:39 | all together and exit. Okay, how virus viral life cycles will look |
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| 09:47 | in large part is what's the nature that genome RNA Dean. Okay. |
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| 09:56 | that's how the middle part here. part may not so much to redo |
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| 10:04 | not so much this part. they look a little different virus the |
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| 10:09 | type. Oftentimes because of what's the type. But the thing is the |
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| 10:15 | . You're gonna get this no matter type you are. Okay, that's |
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| 10:19 | goal. That's how the virus going produce make far apart. So remember |
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| 10:26 | this idea that this right here. . It begins at four ends by |
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| 10:34 | that whole set. Okay, so , okay, I do this in |
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| 10:42 | remember I don't think your book even this. But we just looked at |
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| 10:48 | viruses. Right. And here's your caps that shape. Right. This |
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| 10:54 | sure. Is that geometric shape? , that's very common. But you |
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| 10:59 | have viruses that look like this as where the caption isn't so obvious. |
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| 11:05 | guess it's not like a little house tent around the genome. Right. |
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| 11:09 | just basically intimately math to the Right? So the capsule proteins |
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| 11:15 | Alright. Are sticking to the Right? So these will be all |
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| 11:23 | top of the genome and that's how captured uh, the structure. So |
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| 11:28 | call it nuclear capsule because it's right smack bound to the genome. |
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| 11:34 | so you do have viruses that have coronavirus. Alright. So, |
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| 11:41 | and so I think we ended with and viral. So remember crayons and |
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| 11:47 | are not viruses you might call the like, okay, if you see |
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| 11:53 | statement on exam question says protons and are viruses that's false. So, |
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| 12:03 | , so because they don't have the of the virus, even the virus |
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| 12:06 | pretty simple, viral loads are and don't have those things. Right. |
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| 12:12 | prions are infectious proteins and that's all are. Nothing else associated. And |
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| 12:19 | are infectious RNA. And that's Nothing else is a part of |
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| 12:22 | So I think that catches us up questions about these previous things here. |
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| 12:34 | so let's look at this question So this will take us into kind |
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| 12:47 | classification. And as we're looking at , so in reference to an RNA |
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| 12:52 | , majority of viruses, depending on particular virus type, this genome could |
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| 12:57 | used as a template for what? next is going to be the |
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| 13:04 | Classification table. Um You'll need to that. Okay I'm going to show |
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| 13:11 | just to give you an idea of types of viruses that are out |
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| 13:15 | Um Most common ones, you know the question. Oh alright I just |
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| 13:27 | on let's try it again. Right while I'm blathering extend time so I |
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| 13:38 | . Okay um. Oh okay. . So you don't need to memorize |
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| 13:46 | but just show you some representative Um lots of human viruses. Human |
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| 13:52 | of human diseases are actually in all RNA viruses. So um so this |
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| 14:00 | is gonna relate to the next thing gonna talk about. So counting down |
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| 14:05 | 10 9 five 32. Okay I somebody saw the answers and said okay |
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| 14:18 | go with these people. Okay it d all the above. Okay so |
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| 14:23 | bonus points. Uh um I can't it might work anyway, so |
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| 14:33 | So you have A plus RNA You have a minus R virus. |
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| 14:41 | um the plus RNA virus will fit A. B. Or C. |
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| 14:54 | take our question. Just answer. one of those. What would this |
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| 15:00 | be a template for that is the . You need that. Okay |
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| 15:11 | Yes. So plus that's a plus translations. So this goes back to |
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| 15:18 | . We'll talk about this again a times because it can be taken cars |
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| 15:24 | explanations plus minus. Right. So that what can we do? Pay |
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| 15:29 | . That has its own language? off base pairing three problems. |
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| 15:36 | And the plus minus since Auntie says non coding. Right? Uh And |
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| 15:43 | the plus stand basically any plus strength all this language applies to what you |
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| 15:48 | in A. Or D. A. Right? Because D. |
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| 15:52 | . A. Has plus minus plus RNA and DNA. So yeah it's |
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| 15:57 | so you can have this this this this. Okay in all three of |
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| 16:12 | and all have a plus minus relationship each other. Once coding was non |
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| 16:17 | um the five prime three prime relative each other. Right? So there |
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| 16:24 | a stupid penny um the three prime prime relationship. Right? Five prime |
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| 16:31 | prime. And and the carpentry strand like that. Right? So yeah |
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| 16:39 | but whatever the nucleic acid types coming and all three days are possible, |
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| 16:44 | all have that same kind of terminology use. Okay. And so um |
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| 16:52 | and this I'm bringing this up now as you get to the next table |
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| 16:56 | there are no viruses can be a complicated because of this. Okay so |
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| 17:03 | template for M. RNA synthesis. this guy. Okay minus aren't virus |
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| 17:09 | that genome is a template that you um talking to make A M. |
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| 17:15 | . Okay basically making a plus Okay DNA synthesis that's retrovirus. So |
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| 17:29 | in their own room. They can an RNA genome but they copy into |
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| 17:32 | . N. A. Completely So we'll see that on the next |
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| 17:36 | the next flight. Okay and again go through the plus minus all that |
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| 17:44 | more more than once. Okay so go ahead and flip over here. |
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| 17:50 | um Okay so here again in this . Right so the just think of |
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| 17:57 | plus RNA is the M. It's the same thing happens in you |
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| 18:01 | you undergo express jeans you're taking DNA , your D. N. A |
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| 18:07 | that you copy it um into a . You copy the minus D. |
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| 18:13 | . A. And you got a art. Okay so it's okay this |
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| 18:20 | we're gonna clarify that. Um So this classification there's a question we had |
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| 18:26 | time about what can you use to viruses? All of them are |
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| 18:31 | Right, capsule type, genome type envelope and the envelope legitimate reasons things |
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| 18:39 | can use to classify. But what's used is this Baltimore classification and it |
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| 18:44 | genome? DNA. RNA double strand thread. And what's the rap? |
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| 18:52 | did they get to the M. . N. A. The plus |
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| 18:57 | . Because that's what enables it to which you're gonna need. Right if |
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| 19:03 | gonna assemble into a viral particle. so group one into your D. |
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| 19:08 | . A. That's you know that's we are. Right. So you |
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| 19:13 | should have the greatest familiarity with that that's how we do it. |
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| 19:16 | We don't we have double strand. would be here we have double stranded |
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| 19:20 | of course. But we know how happens. Right? Um transcription to |
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| 19:27 | D. N. A. Into . R. Okay. And so |
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| 19:31 | the single strand D. N. . You first uh make a double |
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| 19:37 | of the form. Okay and then the minus strand into A plus |
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| 19:42 | R. Right? So this is this gets you enable us to make |
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| 19:47 | . Okay. And then uh the groups. So here again is just |
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| 19:53 | example of RNA uh with strand to . So it's RNA RNA. And |
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| 20:01 | know that because remember use right use only RNA. Not in D. |
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| 20:07 | . A. T. I mean a cell thing. Right? So |
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| 20:12 | so the plus strand and what we the coding or the sense strand? |
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| 20:24 | uh mega strand is called the non or template or not? Antisense |
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| 20:32 | Okay. The current trend simply has information to make the. Okay. |
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| 20:40 | so um when we make it we an M. R. A. |
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| 20:45 | here topping that DNA template strength? copying minus the only copy of minus |
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| 20:53 | to make a plus. Right so minus template string enables us to make |
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| 21:01 | plus R. N. A. now contains the essential information to be |
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| 21:06 | into a project. Okay so that's getting to the plus stray. And |
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| 21:12 | with viruses is a big deal because the M. R. That's how |
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| 21:17 | what that's what gets translated minus cannot contain the cooling information. Right? |
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| 21:26 | with the RNA is here they have own enzyme. Okay this RNA dependent |
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| 21:33 | the so you're going well we are but it's D. N. |
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| 21:41 | D. As in dog D. . A. Dependent on limericks. |
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| 21:46 | what we've got viruses uh can't use . Right? They didn't RNA dependent |
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| 21:53 | income because they they have RNA genome that are in a into RNA. |
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| 21:59 | . Us you carry out don't do kinds of things. Okay. Or |
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| 22:03 | carry. Okay that's a strictly viral . So for that reason they must |
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| 22:10 | their own enzyme to do this. um Which is what this R. |
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| 22:15 | . R. P. Is. and so um so you have three |
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| 22:20 | double stranded and single stranded plus and . Okay so when we copy we |
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| 22:30 | the um minor double stranded copy the strand. Okay and we get A |
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| 22:37 | so we can stop right there. the plus strand. Okay Is we |
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| 22:46 | directly we copy that. Okay now gonna tell you why. So you |
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| 22:52 | here right this is already A. R. N. A. |
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| 22:57 | Do we have to go why do have to go any further any |
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| 23:02 | And it does it doesn't stop Okay so go back to what I |
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| 23:09 | earlier. Okay a virus is it attaches to a host cell. |
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| 23:19 | so that plus genome enters to Okay right there. And so is |
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| 23:26 | gonna be enough? Because remember what's end game makes lots of viral |
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| 23:31 | Each has its own genome proteins around . Right so is this entering the |
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| 23:38 | ? Is that enough? No? so yes even though it can and |
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| 23:46 | is translated. Okay it's not enough . And so that's why now unfortunately |
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| 23:55 | easiest solution would be because it gets little complicated. The easy solution would |
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| 24:01 | let's if we need more copies let's that into a plus. Okay can |
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| 24:08 | work? No because you copy a strength it's just a complimentary base |
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| 24:15 | So you can't that doesn't work. it has to go into a minus |
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| 24:21 | mode first. Okay and so this basically what's gonna see underneath here. |
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| 24:29 | so we go from minus our name two plus. Okay. Yes it |
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| 24:36 | make sense but it's just the base rules. That's really what the |
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| 24:40 | This whole thing right, complementary right. We don't make a direct |
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| 24:45 | copy of the strand. Make the cop whether it's a plus to make |
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| 24:50 | minus it might be a copy until . Right. And so that's why |
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| 24:55 | has to go this wrapped. Because again, you think of the |
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| 25:00 | right. They make lots of copies genome stuffed into. Okay, so |
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| 25:08 | of our couples not enough mass Okay, so with the minus RNA |
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| 25:15 | . Okay, they can do it one step. Right here is minus |
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| 25:22 | plus. Okay. And but again you think all right, we're gonna |
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| 25:28 | lots of viral particles. Right. we need to make lots of copies |
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| 25:34 | this because that's the kind of virus is viruses exiting the host has to |
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| 25:40 | a minus genome because that's the kind viruses right way to do. It |
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| 25:46 | to make you have to make more of this as well. Right, |
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| 25:50 | a minus RNA. But this table only really focusing on how do we |
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| 25:56 | to the plus? Okay, here , here, here and here. |
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| 26:02 | I'm just thinking long term. this this minus RNA virus is gonna |
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| 26:06 | to make our particles. So we pockets of the genome as well. |
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| 26:10 | , so again, we'll go through this in uh let me talk about |
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| 26:17 | virus life cycles on right through this thursday. Okay. But I just |
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| 26:27 | to throw it out there now. kind of get it in your get |
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| 26:30 | gear screening on. Right. And um any questions? It's gonna think |
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| 26:42 | it absorb it. I said we'll through this all again on thursday. |
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| 26:48 | so the retroviruses. Okay uh other unique box if you will. |
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| 26:55 | So um they're RNA genome is copied D. A. Okay. And |
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| 27:03 | so the reason for that is why has. What's that we talked about |
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| 27:13 | time. I think HIV is one those types that does what and it |
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| 27:21 | the host. Okay so if you're do that you gotta be D. |
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| 27:27 | . A. Okay. And so actually does that with reverse transcriptase making |
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| 27:33 | minus D. N. A. . And then it actually uses host |
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| 27:37 | polymerase to make the double strand. that's what can get integrated to the |
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| 27:42 | chromosome. So I have to go that step because that's what viruses do |
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| 27:48 | . You're gonna do that the host double stranded DNA. So you've got |
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| 27:53 | make a form of that. So what it does. Okay of course |
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| 27:58 | that it can produce it can use our right to make M. |
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| 28:06 | And A. And then make viral . Okay so um so looking at |
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| 28:14 | types, right again you need to this tape. Okay. But just |
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| 28:19 | you you know diseases that you know diseases that you may be aware of |
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| 28:26 | papilloma virus. Right. Um chicken DNA viruses, single stranded DNA viruses |
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| 28:34 | so much in humans but parvovirus you pets uh cats and dogs. Um |
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| 28:44 | is a bacterial virus here. Um certainly by the RNA viruses. Lots |
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| 28:50 | that are familiar. Okay. Um west Nile is endemic in this part |
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| 28:57 | the country. Um Yellow fever and . If you go uh I've seen |
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| 29:05 | for get vaccinated. You're entering these in latin America especially dengue fever, |
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| 29:12 | fever. Um Certain countries the polio here minus RNA. Or a lot |
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| 29:18 | them. Okay flu Ebola. Uh , mumps, rabies familiar with all |
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| 29:27 | . Right. These are all types coronaviruses. Okay so um retroviruses. |
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| 29:34 | know those types HIV among them. then these groups this group kind of |
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| 29:41 | different these are messy plant plant These para retroviruses. Okay so they |
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| 29:49 | a form of of reverse transcriptase so don't integrate the post genome. Okay |
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| 29:57 | they have this kind of different way replicating. So they produce an |
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| 30:00 | N. A. From their uh . N. A. And then |
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| 30:05 | they basically transcribe. Okay and then is A plus RNA. We can |
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| 30:11 | viral proteins with that. Okay. reproduction replication and but to make its |
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| 30:19 | . N. A. It copies R. N. A. Using |
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| 30:22 | type of birth transcriptase. Okay. make D. N. A. |
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| 30:26 | it's kind of so retro viruses go you see here right. And this |
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| 30:32 | what the pere retrovirus does. What see here. Okay. RNA. |
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| 30:36 | DNA. This one's DNA RNA. to DNA. Retroviruses. Our |
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| 30:41 | And D. N. A. R. N. A. Back |
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| 30:45 | Arnie. Okay so it's just a different. Okay um I'm not sure |
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| 30:50 | they involved that strategy but um these viruses are very used a lot in |
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| 30:58 | in genetic engineering as vehicles to transfer into plants and things for biotechnology purposes |
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| 31:08 | yeah a little bit different in terms how they do their replication. Okay |
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| 31:15 | um. Okay. Alright. Almost done. So the large so I |
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| 31:24 | put this in there to some very . So we saw the size range |
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| 31:31 | viruses 20 to 900 nanometers. So is something that's much larger. Micron |
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| 31:37 | more than that. Okay. These things that typically amoeba large proto |
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| 31:46 | Okay. But they do have their . N. A viruses they do |
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| 31:52 | you know a lot of D. . A. One point over a |
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| 31:56 | base pairs in this one. Um of protein coding genes. Obviously they |
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| 32:01 | have uh rudimentary metabolisms, right? some amino acids a little bit of |
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| 32:09 | and lipid metabolism. I'm not sure extensive it is but certainly more so |
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| 32:14 | your typical virus. Uh Now what these where I come from? It's |
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| 32:21 | that maybe they were a cell at time that's now just lost a lot |
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| 32:26 | these functions. And so it has dependent on affecting another cell type because |
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| 32:32 | that. But are still maybe in to be, you know, maybe |
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| 32:39 | functions will be lost as time goes . I don't know. But it |
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| 32:42 | have been something of that kind of . Okay. Um they're not I |
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| 32:49 | know proportionally how many of these there if there are a significant number but |
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| 32:55 | they are. I think they're found marine environments typically. Uh But then |
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| 33:00 | is it is you see here a that's infecting another virus. Okay, |
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| 33:08 | that's so um the so what can virus beginning from viruses affecting? |
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| 33:18 | obviously it's getting some function. So these large viruses and they do have |
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| 33:25 | metabolisms that viruses don't have. So was kind of a mind bender saying |
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| 33:34 | virus infects another virus. Okay, , it's not something that's very prevalent |
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| 33:39 | you know, it's it's it's out . Okay, so any questions. |
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| 33:48 | So last thing we're gonna talk about is just on equality. So I |
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| 33:53 | earlier last time most of that of viruses were there was no redeeming |
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| 34:02 | Okay, so other than causing Right. Although they do have certainly |
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| 34:06 | use in the lab um as a to transfer genes and whatnot. But |
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| 34:14 | it is shown to be quite um their activity. Okay. So they |
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| 34:25 | marine environments that have been seen affecting and plankton uh that they can control |
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| 34:34 | of marine organisms uh microbes in the . And in doing so when you |
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| 34:43 | control the numbers of populations that allows to kind of thrive as well. |
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| 34:49 | have one dominant species. You will a lot of diversity. Right? |
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| 34:54 | if you kind of viral infections can of affect certain types of lower population |
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| 34:59 | or certain types, you don't get imbalance right? You get more |
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| 35:04 | And they've seen that. Okay. so when they speak of this viral |
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| 35:09 | , right? It's a mechanism really byproduct of infecting cells. Okay. |
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| 35:17 | um of course if you break this you infected cell and cell rises and |
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| 35:22 | really is the nutrients available to others others to use. And that's kind |
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| 35:26 | their contribution here. Is that plus kind of controlling population sizes. So |
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| 35:32 | can see here in this um little here. Right. So we have |
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| 35:38 | specific to bacterial um put synthesizers, environments, those specific for uh uh |
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| 35:48 | like algae and plankton. And so infecting them, the that material becoming |
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| 35:55 | for others to use nutrients, carbon in etcetera. Um but then also |
|
| 36:02 | other micro types to flourish that maybe wouldn't. Okay. And and of |
|
| 36:08 | they don't the viruses themselves don't take population is down to zero. |
|
| 36:16 | Because they're looking all right, So be some types that will be resistant |
|
| 36:20 | the viral infection. Okay, So not talking about taking these things down |
|
| 36:24 | zero. Just reducing numbers. And over time some of the cell types |
|
| 36:29 | resistant to the virus and then it back and forth. And so but |
|
| 36:33 | a way to control numbers. It's way to provide nutrients to the ecosystem |
|
| 36:37 | well. Okay, so um and same thing happens in our gut. |
|
| 36:43 | have viruses that are good that are bacteria in there and and creating the |
|
| 36:50 | pipe controlling population sizes as well. , so so That wraps up part |
|
| 36:59 | . Okay. Life cycles. We'll with bacterial life cycles. And then |
|
| 37:11 | don't understand because in fact the Celtic not as complex right for themselves are |
|
| 37:18 | complex as you carry out. so um any questions before. |
|
| 37:27 | Alright. So this is just a of we talked about before. |
|
| 37:32 | viral life cycles. And so the attachment the genome. So the variations |
|
| 37:44 | didn't step uh so much variation in beginning because recognizing the host and whatever |
|
| 37:52 | type it is about interacting with each on the surface uh genome entry should |
|
| 37:58 | the entire captured which is comedy. animal viruses. Uh bacteria viruses will |
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| 38:03 | need a genome uh for sentences assembly part of the process. Um And |
|
| 38:10 | is then here is where you can exits where you can have the |
|
| 38:15 | Maybe those that have envelopes, envelopes around them or not. Okay so |
|
| 38:20 | go through specific examples of bio types and see some of the variations in |
|
| 38:27 | steps. Okay so uh let's get question here. Okay so this is |
|
| 38:35 | uh fade out fade or bacterial Um And this is asking about uh |
|
| 38:45 | is following is or are not part the life cycle of a lighting |
|
| 38:51 | Little paralytic page. Okay what's So we're gonna have what they were |
|
| 39:01 | about bacteria or viruses there there's different in terms of how they conduct their |
|
| 39:10 | . Lighting is one of those. let's go ahead and go forward from |
|
| 40:10 | 54. Okay. Alright so look the two. Right So E. |
|
| 40:28 | D. Yeah so E. And . Jack. Um So certainly |
|
| 40:36 | B. C. So nature of lighting viruses eventually effect then pretty quickly |
|
| 40:45 | after killing. Uh Of course in middle make lots of viruses. Okay |
|
| 40:53 | let's look at bacteria phage life cycles specific to bacteria. Um Whether you're |
|
| 41:01 | little virus or what we call elissa virus of course ends with recognition. |
|
| 41:09 | . And so bacterial viruses typically have kind of structure. Right? Looks |
|
| 41:14 | the your book calls them tailed Other books call them complex viruses because |
|
| 41:20 | have they have typical capsule structure but they have these other parts to |
|
| 41:25 | Right. And so this actually compresses you see here and in the process |
|
| 41:32 | that D. N. A. bacterial is very common. That that's |
|
| 41:37 | that's the only thing that gets in genome. Everything else stays outside. |
|
| 41:42 | it essentially becomes the protein parts have basically what they call ghosts because they |
|
| 41:48 | have any more just a protein shell husk on the outside of the |
|
| 41:54 | Um Now athletic fage license page. so very versus wait another term for |
|
| 42:06 | pages is what they call virulent. . Try it again here. Same |
|
| 42:19 | virulent. Sorry, try one more . Okay. All right. |
|
| 42:35 | I. R. U. E. N. T. Okay |
|
| 42:38 | there's another name for little viruses um temper. So temper temperate to kind |
|
| 42:44 | go run hot and cold. So stage has a period. It can |
|
| 42:52 | short or long where it's kind of into the host chromosome. Just sitting |
|
| 42:57 | . Okay. Not doing anything. . That's the nature of a license |
|
| 43:02 | stage. Okay so um a pro is what that is called profane. |
|
| 43:12 | what you think, you know, into the host. Okay. And |
|
| 43:17 | gonna show you on the diagram. kind of combines both. So even |
|
| 43:22 | you're a less energetic page and just out eventually you're gonna go into a |
|
| 43:27 | cycle because ultimately you want to make particles. Okay. And the only |
|
| 43:32 | to do that is to go into light cycle. So for less hygienic |
|
| 43:36 | license is part of it as well someone. Right And it's the |
|
| 43:41 | The state of the state of the basically determines how fast it will go |
|
| 43:46 | lighting cycle. Okay, so let's at this here. Um Okay so |
|
| 43:58 | with the little page the try to this move out of the way and |
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| 44:07 | try it this way. There we . Okay so the light cycle um |
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| 44:12 | really gonna comprise this part here. so you see the itself um |
|
| 44:19 | I'm sorry. So then we begin right down the host chromosome. Okay |
|
| 44:28 | so our approach and our proteins are um and so very common in in |
|
| 44:34 | virus life cycle. You'll have viral designated as early and late. So |
|
| 44:41 | genes expressed early in the process. later in the process because there's different |
|
| 44:46 | of the lifestyle. You need some proteins early on kind of maybe it's |
|
| 44:52 | enzyme it needs made for it to . Um Later in the cycle it's |
|
| 44:56 | be proteins needed to kind of assemble exit the cell. So it makes |
|
| 45:01 | to kind of differentiate those in terms very important cycle cycle. So you |
|
| 45:10 | see that the host genome eventually is degraded. Right? And so the |
|
| 45:18 | will actually use those nucleotides for fires use those nuclear times to replicate. |
|
| 45:25 | uh you see the assembly of viral and then um exit. And so |
|
| 45:31 | these lighting stage 52 500 particles per okay can be produced. So it |
|
| 45:39 | very rapidly. Um and so you have E coli suspension add a drop |
|
| 45:49 | lighting faith to it 30 minutes. looks clear as water because the cells |
|
| 45:55 | be obliterated and because as soon as exit of course they want to affect |
|
| 46:01 | and more self. Right? It's happen very quick fashion these bacterial |
|
| 46:06 | Okay. And that's that's it for faith. That's all it does. |
|
| 46:10 | what you see in the dash Okay. Um Nice sergeant will at |
|
| 46:18 | point have that as part of the but it also has this part. |
|
| 46:23 | . And so the genome is basically can see it in purple here. |
|
| 46:32 | integrated into the chromosome. Of course cells unaffected. Right? Obviously it |
|
| 46:39 | affected over here. Right? All resources being used eventually to sell |
|
| 46:44 | It kills it. But over here viral genome profile page right, is |
|
| 46:51 | hitching a ride in the chromosome as cell replicates obviously. So there's |
|
| 46:59 | Okay, so obviously all sorts of , you know now how fast these |
|
| 47:06 | grow? Cells grow so many thousands upon thousands themselves. Now carrying each |
|
| 47:15 | a copy of the genome. so that's like a ticking time |
|
| 47:20 | Right? But at some point it's to enter life cycle. That's the |
|
| 47:26 | way to make our apartments. So it will go back into that |
|
| 47:31 | . It's not just it's everything It's not all at once. But |
|
| 47:36 | potentially all the cells that are carrying profane can go into the you get |
|
| 47:45 | lot of faith produced during this So what is the what is the |
|
| 47:52 | thing that kind of controls? Well I stay in misogyny or do I |
|
| 47:57 | into the light cycle? Okay, um if I go into the logic |
|
| 48:04 | , okay, I'm gonna have I'm be creating these. Let me try |
|
| 48:10 | more time here. Going to be um these right here right by particles |
|
| 48:20 | they will want to go right and more cells. Right? So and |
|
| 48:29 | that's the way for the buyer to itself, right to make lots of |
|
| 48:33 | that can then grow and infect more . Make more virus particles. So |
|
| 48:38 | , so knowing that. Okay, that's the success of that. The |
|
| 48:47 | of that requires what when these dudes out, what's got to be around |
|
| 48:56 | hostess? Of course. Right, it's gonna want to do this |
|
| 49:01 | Right? Going to light cycle when potential is there to have lots of |
|
| 49:06 | soldier. Okay, so knowing Alright uh let's look at this |
|
| 49:14 | Okay, I'll come back to Alright, so I hear that. |
|
| 49:25 | right here there it is. okay, there's the question. So |
|
| 49:33 | look at that. Okay, so would promote that was already on its |
|
| 49:40 | , some caffeine or something here. um let me erase this. Okay |
|
| 49:48 | what with the presence of abundant nutrients , promote less so ginny or promote |
|
| 49:54 | cycle? Always virus the traditional Okay, counting down. Okay, |
|
| 50:46 | down. Okay, so it would um so as I mentioned right if |
|
| 50:55 | guys come out okay and they want seek and make sure lots of pushed |
|
| 51:03 | to infect what's gonna make the host happy and lots of nutrients, lots |
|
| 51:10 | nutrients, lots of growth, lots hosts, correct? Right. So |
|
| 51:17 | abundant nutrients would promote going to the cycle. Okay. Um it's wouldn't |
|
| 51:30 | sense for the virus to go to strategy particles if the chances are one |
|
| 51:37 | the poster. Right? In fact would be the case if there were |
|
| 51:42 | know if there were a lack of that it needs to help the host |
|
| 51:46 | are growing that much. Um And the opportunities if it didn't go too |
|
| 51:52 | excites the virus did. But then something that host host, they're too |
|
| 51:58 | . Okay so better to wait. are, it certainly happens to your |
|
| 52:03 | . Right, when you have a they attract attract bacteria viruses that infect |
|
| 52:09 | allies and other bacteria and they see after you've had a meal that there's |
|
| 52:16 | rise in these um landing page, page is the one that does |
|
| 52:22 | Okay and um there's between meals there's reduction right? The virus is kind |
|
| 52:29 | staying staying in that in that estrogenic . Okay then when you get the |
|
| 52:35 | meal is kind of burst you see more of these um landing page appearing |
|
| 52:40 | they've gone to life cycle with the of nutrients between taken in in the |
|
| 52:46 | to grow. Okay. Um Does make any questions about that? |
|
| 52:53 | Okay so. Okay. Yeah. yeah I think this depends obviously they're |
|
| 53:23 | different different viral types. And so herpes virus yeah it forms the blisters |
|
| 53:31 | you're stressed right? But but I'm sure. Yeah so so it's also |
|
| 53:38 | talking about a that on the skin mucous membranes in your mouth and what |
|
| 53:45 | talking about a completely different environment different . And so I think that's probably |
|
| 53:49 | you want to see differences like So but exactly because even with um |
|
| 53:56 | so because of that some of the virus and we're talking about bacterial growth |
|
| 54:01 | ? Which is going to be different herpes virus affecting your own skin cells |
|
| 54:06 | or epithelial cells whatever are gonna be different scenarios in terms of environment. |
|
| 54:13 | it's good question. Good question. other, Yeah you talk about just |
|
| 54:29 | lighting cycle. So life cycle. So we're talking just about the athletic |
|
| 54:37 | like page. Yeah so with bacteria course you have you have lots of |
|
| 54:56 | they're gonna grow. Okay and so you are a Lysa genic virus. |
|
| 55:03 | ? You have two strategies. Okay can stay as a like login or |
|
| 55:07 | can stay go to lighting cycle. . And so their strategy is tied |
|
| 55:12 | really the state of the host Okay. So if you have abundant |
|
| 55:16 | , the host is gonna grow Crazy. Right? That's gonna be |
|
| 55:19 | trigger for the virus to say okay it's time because I can affect lots |
|
| 55:23 | faith come out lots of hosts to . Right? So if it's uh |
|
| 55:29 | lighting virus and does it doesn't do ? Oh geez okay. I mean |
|
| 55:35 | gonna be um uh yeah that that can be limiting for it too. |
|
| 55:41 | I mean like virus does what it it doesn't have any an alternative. |
|
| 55:45 | in fact it's going to go through cycle and kill the cells. |
|
| 55:50 | Now if if if the population it infecting had lots of nutrients and lots |
|
| 55:56 | cells. Right then it's it's a for it. Right? Because now |
|
| 56:00 | doesn't apply to the infection that has of cells in effect or maybe it |
|
| 56:04 | . Right, so there's been a words to infect. So yeah there's |
|
| 56:09 | both ways but for a lot of that only has only one choice in |
|
| 56:13 | make viruses get out and kill Right. So does that make since |
|
| 56:20 | . All right because the lights a one the land of one has a |
|
| 56:23 | of different options here. Okay. and questions. Okay. So |
|
| 56:32 | 13. Okay. So we got of age. This is a lie |
|
| 56:39 | . We have faith ages which led psychologically. So m. 13 is |
|
| 56:45 | of a third category. Okay. it's what we call a slow |
|
| 56:53 | Okay. And so it's a type can infect doesn't doesn't form a profile |
|
| 57:00 | . So it doesn't it's not a in the sense of a landing |
|
| 57:06 | It doesn't stays outside of And so can see it. Uh here. |
|
| 57:14 | again, it's a filament. This of age. All right. And |
|
| 57:20 | only the genome enters what you see here. Okay. And it stays |
|
| 57:24 | stays in the in the site is doesn't go into the host genome. |
|
| 57:29 | . And so um but as it as you see here. Okay, |
|
| 57:37 | going through the copy genome. McGraw assemble etcetera. Right? And it |
|
| 57:43 | . Alright, so that's okay on host. Remember there's a continuum |
|
| 57:51 | Right minded virus where the host cell not going to survive at all. |
|
| 58:00 | it can be a complete uh my for the varsity is hanging out indefinitely |
|
| 58:07 | soldiers is healthy and happy keep growing in between. So as long as |
|
| 58:14 | not putting too much of a strain the hook. Right? You're doing |
|
| 58:21 | own kind of thing replicating. Doing . No rain the host. So |
|
| 58:25 | live with that. You can actually along and divide function. Okay. |
|
| 58:31 | would be happy if it wasn't in but it can still survive nonetheless. |
|
| 58:36 | obviously it's gonna have a very different time right? With the virus inside |
|
| 58:43 | minus not the virus of course. . So but that's that's what |
|
| 58:49 | 13 does it? Has this slow what slow release means the a life |
|
| 58:55 | that doesn't produce a lot of viral at once. And that doesn't put |
|
| 58:59 | strain on the host. The host still survive and replicate although not as |
|
| 59:05 | as I could without it but nonetheless still replicate. Okay. And so |
|
| 59:11 | what is comparing this with the latex ? Okay. But this M. |
|
| 59:19 | maybe have that the writing phase does was it almost guaranteed of What is |
|
| 59:32 | was M. 13. We must it's not gonna run out of this |
|
| 59:38 | . So it's gonna be So you every generation, right? You see |
|
| 59:44 | cells. Right? And so and be others. But but yeah so |
|
| 59:50 | this this charity means there'll be right? I wouldn't say it is |
|
| 59:55 | , maybe it runs in the area in that kind of blows through all |
|
| 60:00 | they're my host. It has. then what they're gonna do. |
|
| 60:04 | But with M. 13, you less less amount of reproduction. But |
|
| 60:11 | know I always got a host, can be short of infecting a |
|
| 60:14 | Okay so you know different different Okay um Alright so we um so |
|
| 60:27 | course with viral infections and how viruses their thing. Obviously host also has |
|
| 60:35 | right We have mechanisms to counteract a infection. Okay so um and so |
|
| 60:44 | uh and us you know anything that to a viral infection has this as |
|
| 60:53 | option? Okay simply changes occur to surface protein that the virus uses to |
|
| 61:02 | interest. So that's that's a common . Genetic resistance is common among many |
|
| 61:07 | kind of life form that succumbs to infection. So bacteria no different. |
|
| 61:13 | restriction enzymes that's gonna be uniquely Right? You are aware of restriction |
|
| 61:21 | from the D. N. Technology right? How we combine make |
|
| 61:27 | DNA markers using the scissors. The in the nucleus. Is that cleave |
|
| 61:32 | . So uh and of course the used. Common technology all come from |
|
| 61:40 | . Maybe some form of the ones it's those uh they know the bacteria |
|
| 61:47 | use this as their defense against Right? So they modify their own |
|
| 61:52 | modify their own D. N. . Right on on the side of |
|
| 61:56 | scenes typically and by manipulating them and that protects their D. N. |
|
| 62:01 | . From the effects of their nuclear . The viral DNA. Okay. |
|
| 62:08 | one is an example. Right so cut it will recognize the sequence um |
|
| 62:14 | these staggered cuts. Right? So like so okay so it just breaks |
|
| 62:25 | of D. N. A. again if you if you have your |
|
| 62:28 | a zines are methylated right here here the enzyme won't attack it by bacterial |
|
| 62:37 | is protected but the viral DNA is sensitive to it. Okay. Um |
|
| 62:42 | there's hundreds of these things. Different of in the nucleus is now the |
|
| 62:46 | that's I guess the more recent in of discovering as a quasi bacterial immune |
|
| 62:55 | is this CRISPR your knowledge? This probably through more through genetics and using |
|
| 63:02 | as a tool to to fix basically D. N. A. Genes |
|
| 63:09 | humans used of course in that research that purpose. Um But was found |
|
| 63:16 | bacteria as a way in which they kind of remember in a way previous |
|
| 63:24 | infections. Okay. And so CRISPR for these sequences that are put together |
|
| 63:32 | are basically a catalog of prior viral . Okay. So I'm just gonna |
|
| 63:37 | this in kind of bids on the slide here. Okay so we start |
|
| 63:43 | course with viral infection of the And then the first part of the |
|
| 63:49 | is this cast protein this short for . Um so it has a binding |
|
| 63:57 | it can have cleavage ability and so binds to this viral sequence. |
|
| 64:04 | And cleaves out a segment. Okay a segment of nuclear tide clean out |
|
| 64:11 | this viral. Okay then uh that the catalog for that virus. Okay |
|
| 64:24 | gonna store it take that sequence and it in its genome. Okay. |
|
| 64:29 | uh bacterial viruses are generally mostly just . N. A viruses. Right |
|
| 64:37 | we're talking about taking a segment of DNA. Putting in the host |
|
| 64:41 | Okay. And that's what this CRISPR is. It's part of the |
|
| 64:47 | the segment of the host chromosome. it's taking that is occupied by these |
|
| 64:52 | spacer sequences they call them. But of these are Pryor cane originated from |
|
| 65:02 | viral infection. Think of each time virus affects it and it goes through |
|
| 65:10 | process. Okay that it has a a little book on that virus. |
|
| 65:18 | puts it on the shelter library. so that all these are like books |
|
| 65:23 | the previous viral infection. Okay so got that catalog in this part of |
|
| 65:27 | genome. Okay. And so it mobilized right when an infection occurs. |
|
| 65:37 | so we transcribe it into our A. Okay. And then we're |
|
| 65:41 | take those and kind of chop up that they call it. Right process |
|
| 65:48 | transcript into segments. We call the sequences. Okay so again these are |
|
| 65:54 | RNA forms of those DNA segments of viral infections that restored in the |
|
| 66:03 | And now we have RNA forms of little segments. Okay we're going to |
|
| 66:08 | which one of these. Which one these? This one this one this |
|
| 66:14 | is going to be homologous. It's be complementary to the virus that's infecting |
|
| 66:20 | now. Okay and so um if a match right that's what's going |
|
| 66:26 | So you see that cascade protein and with the christmas sequence. We see |
|
| 66:33 | there's a binding take them off. it means that there is that that |
|
| 66:40 | was indeed in this sequence it was by this virus before because it has |
|
| 66:48 | sequence that is complementary to. Okay so the binding leads to basically inactivation |
|
| 66:55 | the virus the viral infection stops. of course the strategy works if like |
|
| 67:02 | said if there was a complimentary segment that region if there wasn't then then |
|
| 67:10 | will likely succumb to that. Um but but that's so it is |
|
| 67:17 | of like a that's what they call , a pseudo quasi new system because |
|
| 67:22 | new system army adapted we have a to we know you know we've been |
|
| 67:29 | by a particular agent before. Right . Your system recognizes it and then |
|
| 67:38 | antibodies to the time is so much way. It's the same thing. |
|
| 67:44 | . Um Any questions? Yeah. . Yeah I think so because it's |
|
| 67:56 | not in terms of the components but in the maybe the function of it |
|
| 68:00 | because you can remember previous viral infection it's stored into that sequence. We |
|
| 68:07 | do the same thing but in a way but it is re recognizing um |
|
| 68:14 | . And so we have a record in our immune system. That's something |
|
| 68:19 | talk about later in the semester. I should go okay of course. |
|
| 68:26 | has a very specificity this caste That's how it's used in human genetics |
|
| 68:34 | kind of fix um mutated genes because able to recognize we can engineer it |
|
| 68:42 | recognize specific sequences. So we can it to recognize a human gene and |
|
| 68:47 | you can use the editing function to of cut out the bad parts put |
|
| 68:52 | place a really good part and hopefully the gene. There's been some uh |
|
| 68:57 | with this. I don't think it's mashed mass market problem. Anyway, |
|
| 69:03 | certainly one of those things uh definitely the future. Um Now you've probably |
|
| 69:10 | about this in genetics class as So here let's look at this |
|
| 69:14 | Okay uh we're gonna switch from transition bacterial viruses, viruses. Okay this |
|
| 69:23 | kind of the intro here. Okay count down from 87. Alright so |
|
| 70:04 | get yeah it's gonna be genome type . RNA. It kind of determines |
|
| 70:10 | we'll go in the cell. Okay um okay so mm bars lifecycle. |
|
| 70:22 | we talked about focus on the Metro is within a hope. How many |
|
| 70:29 | cell types are infected? Right. so we look at broad versus |
|
| 70:33 | Uh this picture showing the binding for virus is on uh preparatory sell these |
|
| 70:43 | are actually present. These are involved binding to cell. So it uses |
|
| 70:49 | to to infect. Um the uh replication cycle? Right? So we |
|
| 70:58 | talked about D. N. A type in large part determines where it |
|
| 71:03 | when it infects a cell. And D. N. A viruses. |
|
| 71:07 | What's so again in the context of viruses And I was strictly talking about |
|
| 71:13 | viruses. Um The where is of they in fact eukaryotic cells and all |
|
| 71:20 | complex structures in eukaryotic cells can be part of the life cycle. So |
|
| 71:26 | the D. N. A virus likely to go? Where the facts |
|
| 71:30 | that like herpes virus or coronavirus? were elected go in the cell because |
|
| 71:38 | it has options right side. Pro cell where there's no structures inside but |
|
| 71:44 | is important eukaryotic cells, right? the nucleus? You're right uh Environment |
|
| 71:54 | . N. A. Is going the nucleus. But what's it going |
|
| 71:56 | find in the so when you replicate ? Right, your chromosomes do this |
|
| 72:04 | the nucleus. Right. They copy . So the DNA proliferates to do |
|
| 72:11 | is in the so that's why DNA typically go there because that's where they'll |
|
| 72:15 | the DNA polymerase to copy their Okay. There's some that do carry |
|
| 72:19 | own but many do not. And very often that's why that's the reason |
|
| 72:24 | going from nucleus is that's what it that we can copy its genome. |
|
| 72:29 | . Um are the viruses we talked this before. Again, that's a |
|
| 72:35 | a virus enzyme, viral enzyme is a host enzyme. Okay, so |
|
| 72:42 | gonna have to have that either express . Uh have the gene for that |
|
| 72:48 | their genome. Okay. Retroviruses we know they're different to integrate into the |
|
| 72:56 | genome. So they're gonna have a transcriptase to do that. So um |
|
| 73:02 | with bacterial viruses, yes, genome everything stays outside. That's pretty much |
|
| 73:10 | standard for bacteria viruses stages. And virus is different strategies. Okay, |
|
| 73:17 | what you see here. So this process is the mechanism by which that |
|
| 73:23 | is released into the cytoplasm. so there's three ways to do |
|
| 73:29 | Okay, so we can basically uh so you see the viral approaches here |
|
| 73:40 | to suffer. And then that envelope uh meld with the self pleasure. |
|
| 73:51 | here is the actual state police. . Um and then genome Captain broken |
|
| 73:59 | GM is released. Right? A or endo zone. Okay, so |
|
| 74:07 | see I believe on a virus and will fuse first commonly use this receptor |
|
| 74:17 | one cholesterol and ourselves enters the same receptor mediated endo psychosis. And so |
|
| 74:24 | virus exploits that receptor and then the forms around it and excuses for the |
|
| 74:33 | own. That's what will digest resolve Captain releasing the genome. Okay. |
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| 74:41 | for a DNA virus like this guy it can again it can form a |
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| 74:48 | around it but instead of being released the outside of it it goes to |
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| 74:55 | nucleus. Right? That's where the first nuclear members. That's typical for |
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| 75:03 | . Okay. Um but methods of confusion at the time of membrane or |
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| 75:12 | information or part of the two common of course, whether releases here or |
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| 75:22 | depends on the virus or DNA Ok folks look finish it there and |
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| 75:29 | it up next time. Thank Yes. So they initiated by without |
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| 76:11 | ? Yeah. Yeah, monday, . But that doesn't work. Let |
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| 76:31 | know if it does work. This a prior infection. Now when it |
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| 77:08 | this process it will have one of segments that will be able to buy |
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| 77:13 | it. And so it does and will inaccurate. So basically this process |
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| 77:19 | successful stops the virus infection, it go any further. Right? It's |
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| 77:28 | . Right, correct. So it's us if we have were vaccinated, |
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| 77:32 | ? And so we get vaccinated with virus. So so that now your |
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| 77:41 | remembers. Right? So next time get infected with that same virus. |
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| 77:49 | , so it's the same thing. question I answered the wrong thing on |
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| 78:01 | last question because I put that in proteins. Okay, so like okay |
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| 78:17 | last part. One more time. it's so I didn't think it was |
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| 78:21 | because I was like oh bacteria just be DNA or RNA as well. |
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| 78:26 | why does the genome type matter? because we're talking about the virus. |
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| 78:34 | the animal virus most bacteria viruses are . N. A. But for |
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| 78:38 | animal viruses the it's um so we're about the type of life cycle. |
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| 78:44 | where does it go in the cell effects. So are the viruses do |
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| 78:48 | typically go to nucleus? They do things outside the nucleus where DNA viruses |
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| 78:52 | go to the nucleus for you Because preliminary typically and things like that. |
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| 78:59 | that's what that's referring to. And what it goes one way or the |
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| 79:02 | is going to be really what kind um does it have? We'll determine |
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| 79:07 | . Okay so that's because we're talking animal viruses and their structures in eukaryotic |
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| 79:13 | . Now animal virus can have different . Unlike bacterial virus. And the |
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| 79:20 | that chooses is mainly the type of , yes. Okay so think of |
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| 79:27 | less of RNA and DNA and more . Yes. Yes. Yes it's |
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| 79:34 | of it's kind of yeah it's about does the virus need. Again there's |
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| 79:39 | exception to these things next time but an RNA virus doesn't need. So |
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| 79:45 | the point of going to the nucleus virus does for the most part so |
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| 79:51 | has to go there. Right so kind of what this is. But |
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| 79:56 | there's there's exceptions because the virus actually . But we're talking about for most |
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| 80:04 | . this is how it works. always gonna be exceptions, fortunately, |
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| 80:10 | , okay, okay, so I 11 30 12 30 monday, |
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| 80:21 | That doesn't |
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