QUESTION 1. Usng he avalable lechnology to syntresize ONA with speoific sequence

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QUESTION 1. Usng he avalable lechnology to syntresize ONA with speoific sequences directly, you made a DNA molecule with the comphte gene sequence or he human insuin gene, including al istrons and esons and inserted it into an expression vectr (including bacterial promoter, Shine-Delgarno sequence, and oe) in order to produce insuin in bacteria. However, you efforts faled to produce fiunctional insulin and even he pre-proinaulin was not made by the bacteria Whst must you Tue' in your vector or the strategy in order to produce functionsl insulin ? Remove the ineran sequences snce bacena cannot carry out spícng ? B Remove the sequences coding for the Srst 24 amino acids in the ,Harmuus snce bacteria sannat pechealy cus the human proein out CProduce the two protein chains (A and Bi separately since bacteris aso canner the proirsulia protein into the heo chains D. Two ofthe above EAllatte above c" a.5 polats QUESTION 2 1. You have isolated a DNA ragmant containing a human gene insolved in lung cancer and are working nserting it into a standard plasmid vector used in molecular cloning which has s mulsiple cloning site in he lacz gane Arter cuting the DNA ragmant and the wector wth the same resniction enzyme and igating the out DNA, you ransfom bacternia with hopefully the recombinant vector. How do you expect t dentify bacteria wih the recombinant vector? ? A. Bacens wth he fecembnant vector wil be resistant to antitoonc treament and form blue conn-s in the presecsor X-gal r B. Bacteria with the recombnant veclor wi be resistant to antibiotc treatment and form white c asena wththe recombinar t vector w l te se stie toattiotic treatment (die) and to m whte D.Bscteris with the recombinant vector wil be senstive to antibiobc trestment (diej and form blue oolonies in the presenceof X-gal oolonies in the presence of X-gai oolonias in the presence of X-gal ? r E None of the aboe .5 pelats QUESTION S 1. In your forward genesic study, you hawe identfied two mutants with non-iethal defects in your phenotype of interest. You perfomed a complementsson test to see if the mutations are in the same gane or two different genes but the offspringsfrom the test die eary on in embryogenesis. You condude then that (select the most Lkely expianation) r A the mutations are in the same gene r 8 the mutatons are in teo diferent genes C the mutsions must cause frameshifts in order to produce the lathal phenotype C D.Two of the above r E Cant tal from the information given 0.5 points QUESTION 4 L. At the end of a forward genetic study, you end up with. 5 ?alist ofcandidateients involved inyour phentype of internt. H a list ofphenotypes assockated with mutations in your genes of isterest with mutations in your genes of isterest &cones; of your penes afinteres r D Two of the above E All nf the abave

Explanation / Answer

Q1. Remove the introns since the bacteria cannot remove introns. Bacteria donot have repetitive genome which is foun in introns onnly unique genes, thus they do not possess any mechanism to remove them

Q2. the recombinant vector will be carrying resistance to a particular antibiotic so the transformed cells will be resistant to the antibiotic supplied in the agat medium and thus they will grow. However, the white colony will carry the gene of interest as the gene of interest will disrupt the lac gene. The blue colonies will have an intact lac gene forming functional Beta galactosidase which will hydrolyze X gal into blue product and will lack the gene of interest.

Q3. cannot be determined

Q4. list of phenotypesassociated with mutations in your gene of interest

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