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Cirele " ALULSplall tbe correct answer (? pts each) FExampks € of reporet [ lacz genes afe I, luc IV_bla All of the - following- are INCORRECT EXCEPT? al...

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Cirele " ALULSplall tbe correct answer (? pts each) FExampks € of reporet [ lacz genes afe I, luc IV_bla All of the - following- are INCORRECT EXCEPT? all are inconect cnc [nc omeci (c) L, Hand III only 1 RIn recombinant DNA methods, the term Uctorcan refer 5 (2) the enzyme that cuts DNA into restriction fragments plasmid used transler DNA into , living = cell the sticky end of a DNA fragment a DNA probe used Identily particular gene(d) IL, IIl and IV onlyR*Animals unacs readily manipulat

Cirele " ALULSplall tbe correct answer (? pts each) FExampks € of reporet [ lacz genes afe I, luc IV_bla All of the - following- are INCORRECT EXCEPT? all are inconect cnc [nc omeci (c) L, Hand III only 1 RIn recombinant DNA methods, the term Uctorcan refer 5 (2) the enzyme that cuts DNA into restriction fragments plasmid used transler DNA into , living = cell the sticky end of a DNA fragment a DNA probe used Identily particular gene (d) IL, IIl and IV only R*Animals unacs readily manipulated by genetic engineerng than are plants because animal genes contain only exons animal cells have larger nuclei animal cells are totipotent All of the following WNCORRECT EXCEPT? all are inconect (6) none Incomeci (e) Tand Il only and II only R#The complete and final Arabidopsis genome sequenced (4) by sequencing each chromosome, one at 4 time (6) using DNA fingerprinting looking for overlapping regions betwcen sequenced Bcne (rgments using open Teading frames The future of molecular biology will involve USIg all the omics bioinformatics understand the organism pharmacogenomics systems biology- using artificial DNA recreate new organisms Synthetic biology using SNPs to create medicine following are CORRECT EXCEPT? AIl of the Iand I[ only (b) H and III only (c) all are correct none conecl RRR*An infectious agent that has nucleic acids CANNOT be a bacienum prion Mins bacteriophage RRRIf the _ content 0f dsDNA 2594, the U content MUST also be (6) 50%0 2590 there no answer



Answers

Describe the process of molecular cloning.
a. The foreign DNA and plasmid are cut with the same restriction enzyme and DNA is inserted within the lacZ gene, whose product metabolizes lactose. The foreign DNA and vector are allowed to anneal. The vector is transferred to a bacterial host that is ampicillin sensitive and those with a disrupted lacZ gene show inability to metabolize X-gal.
b. The foreign DNA and plasmid are denatured using high heat, and DNA is inserted within the lacZ gene, whose product metabolizes glucose. The foreign DNA and vector are allowed to anneal. The vector is transferred to a bacterial host that is ampicillin sensitive and disrupted lacZ gene will metabolize X-gal
c. The foreign DNA and plasmid are cut with the same restriction enzyme and DNA is inserted randomly in the plasmid. The foreign DNA and vector are allowed to anneal. The vector is transferred to a bacterial host that is ampicillin sensitive and the disrupted lacZ gene shows inability to synthesize X-gal.
d. The foreign DNA and plasmid are cut with the same restriction enzyme and DNA is inserted within the lacZ gene, whose product metabolizes lactose. The foreign DNA and vector are allowed to anneal. The vector is transformed into a viral host that is ampicillin sensitive and the disrupted lacZ gene show inability to synthesize X-gal.

This question is making sure you understand restriction enzymes, how they cut and how they're sticky or blunt ends will react with different portions of DNA. Create a new hours double strand set of DNA. So part A is asking us. But a Nico are one enzyme cut would look like so in one of the tables in the textbook. It gives us the sequence for ICO, our wine. It's going to look for a G, A, A, T T C and its complementary strand of C T T A A. G. And here the ICO are one enzyme is going to cut along this pathway. It's going to separate the two ends and create two fragments of DNA along this point. So for part A, we're going to have a long strand of DNA mixed with It's a and its complementary par shin of the strand with a T. T. A sticky end, and this is going to be separated from the rest of the fragment of the single strand of DNA that we just cut you have. It's three prime and five prime, so these are the kind of things you'd see. Just remember, we have a longer strand of DNA continuing out along both sides. But this is the site we're looking at. That was just cut for B. It's just asking us to attach a nucleotide triphosphate to the sticky ends. So we're just going to attach the complementary bases for each of these fragments. So here on the three prime Strand, we have our t t a. So we're going to attach our complimentary A T T. And then on the second fragment we're going to attach are complementary pairs like ways. So we have a okay to to. So these are new fragments we've created with blunt ends. Now, since we've added our nuclear tied try phosphates to each of those for part C, it's just asking us to attach kind of these blunt ends together to create a single strand of DNA has just been reassembled. So here we'll have our G A T T fragment, and it's going to attach and, like, eight to our A a two TC and Mike wise on the bottom. That's May. So here we have our new DNA strand when are previously made and nucleotide triphosphate added strands have been connected. Just remember, we have our five, prime to three Prime and the complementary strand as well. For Part D, it's asking us to take our original sticky and fragments and to digest any single strand of DNA that's present. So here we're going tohave are sticky ends made with just an original eco r one cut and we're going to digest and remove our UNP aired nucleotides. So in the end, we're going to get very short fragments of just a G here, iguana zine and a cytosine and the other fragment with the site is seen on top and a quantity nonviolent. So here's our new strand we've created. If we're going to digest those UNP aired nucleotides from the sticky ends for party, it's going to ask what happens if we stick our original eco r one cut to what we've created here with just our single nuclear tides. So if you imagine we're going to take our sticky and pears Ah, and we're going to attach this three prime to five prime to our single nucleotide fragment of just the guanine from part D. So here we're just taking this single nucleotide fragment of the five prime and attaching it to the ico are one cut here. Likewise on top. It's just gonna be the complementary, but with this cytosine attached or with a guanine, because we're attaching our a a TTC to our single nuclear Tiggy from before. So for this instance, we are attaching to this five prime to three prime end here, and we are attaching our A, t, t c and G fragment. So this is what they're looking for, which is, ironically, just the ICO are one restriction site as well. Basically, what we've done is we've cut our original strand of DNA removed, are single nucleotides and added them back again for part F. There's actually a misprint in the book. Um, we're looking for the restriction kind of product if we're using a P V U two enzyme. Yeah, So if you notice in the book, the ICO are five. Restriction site is listed as the same as the PV You one PV YouTube PTU to actually has a different restriction site of C A, G, C T G, and its complementary G T c g A C. And it's going to create blunt ends by cutting straight through the center. Try that and read So for going for a PV you to cut, we're going to make the fragments C a g by prime three prime GTC and the fragments ctg g a c So this is what they're looking for for part G. We're going to attach this fragment we've made and add it to the blunt ends that we made in part B. So we're going to add our TV You to to this section here. So, Fergie, we take our five prime three prime initial strands C G G c, and we're going to add it to those blunt ends of our five prime to three Prime and our three prime five Prime. We're going to make this new strand here. So we've just attached are blunt ends together along this point, and this is our new strand with me for part h. They're asking us to apply this knowledge to kind of make us try and get a different plasmid structure where it would no longer be digested by an eco r one site, but it will have a bam h one site. So from what we've learned in the previous questions, what we could dio is we could take our initial Iko are one G A, T T C and its complementary strand Cut it with the ICO are one. And we can either remove this single strand DNA using a different enzyme, or we can fill in here using our nuclear tied complementary pairs. And then we can just add additional nuclear tights on here That would give us a bam H one site as long as we don't begin with C and G along this next nuclear tied we add on. If we add in this CG pair equal are one would still be able to cut it. But if we add, say A and A and A T here equal are one will no longer recognize this sequence and it won't be able to cut now it takes a lot of time and scientific error. Um, restriction enzyme aren't perfect, and they don't give you 100% outcome and yield. So a better way to do this is to create our own DNA strand. But they have listed is we're going to make this artificial strand here and its complement so feel notice. What we're now including in here is our a t T See are these editions to this nucleotide sequence. So basically, this sequence is made by cutting it away with a Nico are one enzyme on both ends and you'll notice the Bam H one restriction enzyme of G A, T, C C, and its complement is located right here in the center. So by artificially creating this sequence of DNA, we have an opportunity to attach it to a Nico are one enzyme site, and it automatically has a bam H one enzyme site included. So, after we've introduced equal art one toe wherever this is attached, we can now make a new cut down the center of this sequence with a bam H one cut. So if you look in this previous example, if we're going to add DNT piece here, we create a blunt end, and blunt ends aren't very good at connecting up ends of DNA. Um, it's easier with sticky ends because complementary pairs like to stick and fix along to make double stranded DNA. Rather than bear up to blunt ends, there's just more free energy available and sticky ends make the reaction more spontaneous and more likely to occur and give a stronger hold rather than just attaching across the DNA rather than across the compliments. So for H, this is our better option. Two years, due to its advantages of sticky ends and its utility of both ends for a Nico are one enzyme and the inclusion of the Bam H one site straight through the center finally per part II. They're asking us to create different strands that are compatible with Iko are one and or PST one. So just as a reminder, all right, the equal are one cut site and the PST one cut site. So for equal are one It's the G A, T T c. And for PST wine, we have CTG, C, A G, and our method of cutting creating sticky ends. So what we want to do is create a sequence where you have hey is both being c N t. Do you is neither for being able to do an ICO are one site and for C, just a pst one site. So, for a what we can do is we can make an eco r one site along the far ends of the strand of DNA, and we can make the PST one sites closer into the center. So if we start with this sequence of DNA a long way, we're cutting. So here we'll have a a a T T. C bound to its G here would be the ICO, our wine cut site in the center. We could use whatever nucleotide sequence we want. It doesn't matter, because we're not going to cut along these lines. And along the right side, we'll have our PST one cut site. So here is our c g C. A sequence. If we wanna have unequal are one on Lee, What we're going to do is we can just change this sequence. This base pair here MPs Taiwan will no longer recognize our strand so we can have a a t t c whatever we want in the center. And then we could just add in a guanine here instead on create are complementary sequence in the middle. So here we stole of our eco r one cut site. However, on this and this sequence is not recognized by any of the enzymes. So for part c, we do just the opposite and change this base pair here and leave this original water alone. So here we'll put in a guanine instead of that side is seen. Keep our cytosine on this strand and drawn are complementary. So here we have our PST one cut site and no cuts site over here. Lastly, for our neither we can change both of these sites so that neither iko are one or P s t one will recognize them. We have a a t t. G. So there's no cut and G T g c A. So there's no cut, so you'll notice neither this strand or this strand is recognized by either equal are one or P s t. One. So there will be no cut along these points. So this sums up how we've located where to cut along Our enzyme restriction site had to attach blunt and sticky ends and how to form different pairs so that were either removing the restriction cut site or adding it into it. And this is what this question is looking to address

Let's talk about Port A Her a Onley clone a not BC or Dean is activated by F gs. So which indicates to us that the DNA binding site, which is Onley located in Clone A, is located somewhere each Wayne, the three prime end of Exxon, one of E one s and the five prime end of Exxon three e three. That's for be part. We can see that cortisol is responsible for repressing transcription and clones A and B transcription Olbricht. Repression is not occurring because of cortisol. Includes scene. And so the DNA binding site must be located in the three prime region of E three s or in the flanks of the three prime int. And finally see part clones C and D or activated by E. T and not Clem being so. What this tells us is that the DNA side involved must be located to the three prime side uh, e three Jeanne. Oops, let's talk parentheses around the three. You need to be around the G. What's just trying? Get in here. There we go. Or again in the three prime sequences in the flights of the global gene

But this question. We have a really long list of be important sums in the straps of as and descriptions to match. So I'm gonna get through descriptions and match them to be. It's, um barracks, describing a intervening sequences found in many eukaryotic treats that these are the entrance is what we want to spice up before. Translation over. Then we have be conserved. Nucleotide sequence in oh, character promoters involves initiation conscription. It's going to one of our sequences on this one is Vita sequence here must be have C Small army molecule was in meat nuclear about Karadzic's owes its components of spices own, displacing the excision of insurance. Of course, this is Snr have d A sequence in the non templates around matters of ICO. Life facilitates from applies, so this is never sequences, sweetheart. Top secret E. Coli is a very important model organism worth remembering. Lots about E, but only memories. But cancel eyes is the synthesis of all our days. Except for the five s subpoena. We've got three irony primaries. Is it one butts? Does most of the represent Laurene? It's only been raised. Then we have if he subunits of pro Carol tick arm in eight primaries that is responsible for the initiation of transcription. That's promotions. This is be Sigma factor. Yes, and E. Coli promoter sequence. Okay, go on the sequence. 35 nucleotides upstream initiation sites. A recognition sites for significant. So this recognition for Sima fax it iss TJ this the lots of these, but it covers all of the important sums. Make the irony play Marie's to left that capitalizes for synthesis of tea on a on a small nuclear. So for this one, h he's going to be are in a play Marie's three. No. Yes, we have I a poly Adina seen fracked, but it's added to be three prime end of Mostar Aquatic. So this is the pulley A to is well described by his name. Okay, we are in a primaries but transcribes nuclear genes. So this is the only one left arm in a primaries to So this makes the Aymara nays and we have K a conserved sequence and the sequence in the non template strand of oh, character promoters. 80 nucleotides upstream from the transcription start sites. Schools be this one last one hit. Indeed. This is a sequence phone them. We have L segments of their character, Teen corresponding to sequences and we finally processed are a transcript. So after spicing, this is what is left. And that's the Exxon's. Yeah, and finally we have and a population approaching transcripts, too many clips of their parents. So and this is it's true genius are in a that Sullivan.

So here we're continuing on with our work related to DNA, and we're gonna be taking about that restriction enzymes. So in the fast pothead of the types of plasmids that could be found with the T r a r with superscript on includes the original intact plasma with the eukaryotic gene fragment on like a tick plasmas with other plasma. It's instead of foreign DNA fragments, so moving on to the next part. So given the data that we have, so the lane wanted to have digested plasmids with a single eukaryotic gene fragment in different orientations. So after digestion, delaying one and two hard only to fragments with different molecular weights, whereas Lane three has a digestive plasmid with two eukaryotic gene fragments, so it yields three DNA fragments instead of two because it has to e c o r I cleavage sites


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