(by senthil.impres from googlemail.com)
Wed Feb 17 08:53:37 EST 2010
I have a question on Em7 promoter. I am working on a BAC construct at the
moment which i would use for generating a transgenic line. I have already
made the targeting vector and now I need to recombine it with the BAC. But
the problem is, I donot have a marker to screen for the positives, i.e, the
BAC clones that carry the targeting vector in it. I dont want to use lox or
Frt system, So I have decided to incorporate a EM7- neo cassette driven by
a E.coli promoter EM7 at the 3' end of the gene of interest thats present
in the targeting vector. This way I could screen for the BAC clones that
have kanamycin resistance (the positives) in petriplates and comfortably
forget about it when I am making the transgenic lines , hoping that
kanamycin wont be expressed in the mammalian cells.
Any idea on these?, do anyone know how strict is this EM7 driven expression?
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