Question

In a hypothetical scenario. You wake up one morning to find your hah has suddenly grown. YOU have been supplementing your diet with a strange now fungus purchased at the local farmer's market. You take samples of the fungus to your lab and you find that this fungus does indeed make a protein (the her E protein) that stimulates hair growth. Yon con struct a fungal genomic DNA library in E. Coli with the hope of cloning the hart gene. If you succeed you will be a billionaire! You obtain genomic DNA from the fungus digest it with a restriction enzyme, and clone it into a cloning vector. What features must be present on your plasmid that will allem you to use it as a cloning vector? How do you distinguish bacterial cells that canny a vector from those that do not? You could screen your library by hybridization with a probe. What information would you need to do this screen? You screen your library by hybridization with a probe and identify a recombinant vector that contains the complete haiE gene. In the meantime, you have developed an antibody to the haiE protein You use cells carrying the vector that contains the complete harE gene to test how well this antibody reacts with the haiE protein. You do the experiment and find that the antibody does not react with the cells containing the recombinant vector. Does this results indicate that your antibody does not react to the harE protein? Explain.

Answer 1

A. Feature-length cloning site, appropriate marker, genes to replicate in E.coli, like origin of replication.

B. There are few screeing methods with which selection can be done . For example blue white screening method. And ampicillin resistance.

C. probes are oligonucleotide sequences. If the gene sequence is known primes can e made for the specific site and a probe can thus be made. Probe must have an marker to check the fluorescece pattern upon hybridization.

D. as antibody will react to the expressed protein and not the recombination vector it is not giving expression regarding the same.