发信人: Ras (很颓的猪。), 信区: Biology
标 题: Re: KO
发信站: The unknown SPACE (Fri Jun 23 11:07:31 2000), 转信

for stable transfection: u include a selective marker (such as G418,
puromycin) in the DNA construct will transfect cells with. Then by
including the antibiotics (G18, puro...) in the media, you will be
able to seletively grow up the cells trasfected with your construct, while
killing the untrasfected ones.

For transient transfection: you don't have such selective markers in
the constracuts. You will always grow cells in regular media. Since it
is impossible to achieve 100% transfection, there is always a good portions
of cells will not be trasfected. After a few generation, those untransfected
cells most likely will out-grow the transfected cells since they usually
have better growth potential (in most cases, transfection brings some
defects to normal cellular activities).

I think your understanding on transgenic and knockout is pretty much
correct, although I am not sure about the exactly right answer.



【 在 netbuggie (我爱QUAKE3D) 的大作中提到: 】
: is there anybody here doing knockout? have a question.
: what is the difference between transgenic and knockout?
: i think transgenic is mediated through microinjection of virus-based
: cDNA, while knockout requires homo.recombination in ES cells.
: also, btw, transgenic is like stable transfection, rt? i do not
: understand the diff. b/w transient vs. stable transfection. can
: any prof. guy give me an explaination?


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