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Consider the figure below and answer the question that follows. prom vv the plasmid shown is similar to what was used to produce the 1 sars-cov2 mrna vaccine.
Consider the figure below and answer the question that follows. prom vv The plasmid shown is similar to what was used to produce the 1 SARS-Cov2 mRNA vaccine. You are tasked to insert the sequence of the immunogen. The restriction map of the immunogen is shown. 1. Your friend who is still in 1 year indicates that your immunogen doesn't have restriction sites on the extreme ends (right at the beginning and right at the end). This means that there is noD way to digest it without losing some of the immunogen sequence. Do you agree that there is no other way but to lose some of the sequence Why? 2. Your colleague advises you to use one enzyme to digest your immunogen and plasmid for future ligation Your Bio206 friend advises you to use 2. Which option will you choose? Why? 3. Which enzymets) will you choose? Why? 4What enzymels) will yotu use to digest each of the 5' and 3 ends of the immunogen? Why?
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Krishan k jakhad![best-answer]()
1. Yes i agree there no restriction sites at extreme ends so in such cases we can use adapter which are chemically synthesized ss or ds- oligonucleotides that can be easily ligated to ends (linkers) to avoid any sequence loss. see attached diagram 2. restriction enzyme Not I cuts DNA at the sequence GC/GGCCGC. okay we can opt for it. 3. already Not1 site is there at plasmid and 5end of immunogen only we have to ligate it at 3' end linker. 4. always we hv multiple options #1we can use only Not1 if using linker #2 we can ligate adapter on both ends and can use Hind3 #3 we can use a combination of Not1 and Hae3. see second diagram bcz there restriction site have nucleotides in common. @ immunogen is a specific type of antigen that is able to elicit an immune response. Antibody development is dependent on a humoral immune response mediated by immune cells recognizing a molecule as being foreign.