Let's assume that I fould a valuable gene fragment (ex. DNA sequence homology with one of the human myosin II genes)
1. Before I proceed for therapeutic development, How would I know if this gene is actually expressed in humans? What method?
2. Now let's assume it is expressed in human. I wish to express the cDNA corresponding to this gene in order to determine its function. How would you construct an expression vector? Would you use the genomic fragment or a cDNA?
3. How would you transfect your expression plasmid into cells, and what type(s) of cell might you use?
4. How would you confirm expression of your construct and encoded protein? (functions and methods)