I have finally isolated a small amount of attractase, an enzyme producing a powe

Question

I have finally isolated a small amount of attractase, an enzyme producing a powerful human pheromone from hair samples obtained from Daniel Craig. My ex-BFF Tom Cruise is having a hard time getting any of the ladies to like him. He has contacted me and asked me to produce attractase, so that he may become more attractive. I obtained a complete genomic clone of the attractase gene, connected it to a strong bacterial promoter on an expression plasmid, and introduced the plasmid into E. coli cells. Tom and I are devastated to find that no attractase was produced in the cells. (Total 2.5 points) a. What is a likely explanation for the failure? b. How would you modify the experiment to increase my likelihood of success?

Explanation / Answer

1. "complete genomic clone" won't work as e.coli doesn't have splicing mechanism it won't get the exons from the full sequence. 2. I would use full exon sequence to clone to pichia pastoris. e.coli doesnt have all post translational modifications of eukaryote which may be needed for proper secondary structure of the protein.For that using other expression system would be better.

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