You are a researcher at a small biotech company and your company has just obtain
ID: 260218 • Letter: Y
Question
You are a researcher at a small biotech company and your company has just obtained the license for use of a human GENOMIC DNA fragment putatively encoding a potentially novel protein, which is thought to regulate p53, the known tumor supressor protein. The scientists who originally cloned this GENE fragment HDM5 "claim" that HDM5 shares 90% DNA sequence homology with one of the HDM2 genes (refer to the review Levine & Oren, 2009). They propose that HDM5 may have HDM2-like properties and may be involved in regulating cell proliferation, and thus a good target to potentially develop as a cancer therapy. Your company has asked you to characterize the gene and gene products, as well as to provide an opinion as to its potential human therapeutic uses.
After convincing yourself that this gene is expressed in humans, you now wish to determine the potential function of this protein by preparing cells in culture which express the protein.
f. How would you confirm expression of your cDNA? Explain.
Explanation / Answer
To confirm the synthesis of cDNA you may run the synthesized cDNA on RT-PCR. Make primers specific for the segment of the particular gene. If you see an increase in the amount of the cDNA then it was synthesized successfully.
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