You are working for a pharmaceutical company are are tasked with creating E. Coli that express an influenza antigen (a protein that is recognized by our immune system) for treatment of the influenza virus. The cDNA of the antigen is available in a carrier vector (pCARRY) that contains a kanamycin resistance gene as shown on the left below. In order to get E. Coli to express the antigen, you need to clone the gene for the influenza antigen into an expression vector that contains an E. Coli-specific promoter. You obtain such an expression vector, which contains an ampicillin resistance gene as shown on the right below (pBACTERIA) a. An end generated by digestion with BamH1 can be ligated to an end generated by digestion with BlgII. Why is this possible? b. To clone the gene for the influenza antigen from pCARRY into pBACTERIA plasmid: i. What restriction enzyme should you use to digest pCARRY? ii. What restriction enzyme should you use to digest pBACTERIA? c. After you ligate the products of your restriction enzyme digests in part b together, and transform E. Coli with the vector obtained, what antibiotic(s) should you culture your E.Coli in to select for the transformed bacterial cells? (circle one) Amplicillin Kanamycin ampicillin and kanamycin
Gene Interactions
When the expression of a single trait is influenced by two or more different non-allelic genes, it is termed as genetic interaction. According to Mendel's law of inheritance, each gene functions in its own way and does not depend on the function of another gene, i.e., a single gene controls each of seven characteristics considered, but the complex contribution of many different genes determine many traits of an organism.
Gene Expression
Gene expression is a process by which the instructions present in deoxyribonucleic acid (DNA) are converted into useful molecules such as proteins, and functional messenger ribonucleic (mRNA) molecules in the case of non-protein-coding genes.
You are working for a pharmaceutical company are are tasked with creating E. Coli that express an influenza antigen (a protein that is recognized by our immune system) for
treatment of the influenza virus. The cDNA of the antigen is available in a carrier vector
(pCARRY) that contains a kanamycin resistance gene as shown on the left below. In
order to get E. Coli to express the antigen, you need to clone the gene for the influenza
antigen into an expression vector that contains an E. Coli-specific promoter. You obtain such an expression vector, which contains an ampicillin resistance gene as shown on the right below (pBACTERIA)
a. An end generated by digestion with BamH1 can be ligated to an end generated by digestion with BlgII. Why is this possible?
b. To clone the gene for the influenza antigen from pCARRY into pBACTERIA plasmid:
i. What restriction enzyme should you use to digest pCARRY?
ii. What restriction enzyme should you use to digest pBACTERIA?
c. After you ligate the products of your restriction enzyme digests in part b together, and transform E. Coli with the vector obtained, what antibiotic(s) should you culture your E.Coli in to select for the transformed bacterial cells? (circle one)
Amplicillin Kanamycin ampicillin and kanamycin
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