I am doing a knockin project. I have done constructs which contain Neo and
HSV-TK for selection in ES cells.
How do I know Neo and HSV-TK in my plasmid work? Specificly,
Do Neo and HSV-TK genes in my plasmids express their proteins in E.Coli? I
tested in this way: grow 4 tubes
of E.Coli, when adding G418, E.Coli can't grow; When adding gancyclovir, E.
Cori grow well. That means Neo
and HSV-TK genes of my plasmids don't express in E.Coli, right? Then why
do Neo and HSV-TK express in
ES cells? I am not sure now. I am treating ES cells with G418 and
gancyclovir.
The sequence of Neo and HSV-TK is right.
thanks