发信人: whlist (windy), 信区: Biology
标 题: Re: Help! Lentiviral expression system
发信站: Unknown Space - 未名空间 (Wed Jun 2 09:43:29 2004) WWW-POST

【 在 tangerman (tanger) 的大作中提到: 】
: Is there anyone who has ever used lentiviral expression system?I got some
: problem.
: 1. After recombination of my entry vector with the destination vector, I
: transformed the expression vector in Top10 or stab (provided by company). I
: got nicked DNA. How could that happen?
: 2. I went ahead and made virus stock and transduced into my mammaliar cells.
: After antibiotic selection, some cells survived. but I couldn't detect my
: protein of interest. How to explain this? What I can do with it?
:
: Thanks a lot
:
:

If things happened like that, why not design a control such as GFP insterted
into your vector, so it is easy to check the infection efficiency. It is not
surprised some cells survived after drug selection, there is why you need to
confirm the ectopic expression of your targer gene in stable cell line by WB
or other ways.


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