rtta/tta primer question

[chef]

hey guys, there's gotta be someone here working w/ TRE/tta inducible mice - i need to know whether i can use rtta primers to genotype for tta, or vice versa?

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[BDavis]

hey guys, there's gotta be someone here working w/ TRE/tta inducible mice - i need to know whether i can use rtta primers to genotype for tta, or vice versa?

We have are in the process of generating inducible mice, but we first started with inducible stables cell lines. For our vectors (Clontech), the rtTA construct we have has a slightly different DNA binding domain from the tta construct (I think 4 point mutations). We have not made it to the mice yet so I can't directly answer your question, but maybe this information will help. Sorry.

P.S. The VP16 antibody from Clontech picks up extra bands on a Western (just in case you are planning to use it; you might as well buy a Santa Cruz Ab )

 

[xanthines]

I am also in the cell culture stage, and you are correct. There is only a 4 amino acid difference between the tta and rtta fusion proteins. That's 12bp, so I guess it could be different enough that the prmers won't work. Just not sure.

Anyways, primers are pretty cheap. If you need some, just buy them. I think they cost like $12.

-X

We have are in the process of generating inducible mice, but we first started with inducible stables cell lines. For our vectors (Clontech), the rtTA construct we have has a slightly different DNA binding domain from the tta construct (I think 4 point mutations). We have not made it to the mice yet so I can't directly answer your question, but maybe this information will help. Sorry.

P.S. The VP16 antibody from Clontech picks up extra bands on a Western (just in case you are planning to use it; you might as well buy a Santa Cruz Ab )