My exam, pls answer - (Aug/21/2010 )
You want to clone gene X which encodes enzyme X from bacterium into E.coli, suppose there are some sequences of the same gene from
other organisms available in the database but with a very low DNA-homology (40%) with each other.
a) What will you do to clone DNA fragments putative harboring geneX?
How will you select E.coli transformant harboring geneX? Suppose there was not suitable protocol for assaying enzymeX.
c) If you want to know whether geneX is expressed in E.coli or not. What will you do?
d) If geneX is not expressed in E.coli, how will you do to get the geneX to express in E.coli transformant?
e) How will you get purified enzyme from the E.coli transformant?