C***Y 发帖数: 61 | 1 I am trying to generate cell line with inducible knockdown. I used Tet-pLKO
-puro, in which a puromycin resistant gene is placed downstream of pGK-TetR-
IRES. The problem I am having now is that the expression level of puromycin
resistant gene was so low that all the cells were killed by puromycin. I
heard that the single vector Tet-induicible system selling by Clontech is
not very good either. Have anyone found any system satisfactory?
Thanks. | b**********8 发帖数: 349 | 2 Why did you say the single vector Tet-induicible system selling by Clontech
is not very good? In contrast, I read many papers in which the authors used
the syetem to devolope inducible kd stable cell line and conduct in vivo&
vitro study. Below are some of them,
1)THE JOURNAL OF BIOLOGICAL CHEMISTRY VOL. 283, NO. 29, pp. 20383–20396,
July 18, 2008
2)J Clin Invest. 2010;120(12):4289–4302. doi:10.1172/JCI42015. | C***Y 发帖数: 61 | 3 Thanks for the response. I found the information here.
http://www.protocol-online.org/biology-forums-2/posts/7444.html
Clontech
used
【在 b**********8 的大作中提到】 : Why did you say the single vector Tet-induicible system selling by Clontech : is not very good? In contrast, I read many papers in which the authors used : the syetem to devolope inducible kd stable cell line and conduct in vivo& : vitro study. Below are some of them, : 1)THE JOURNAL OF BIOLOGICAL CHEMISTRY VOL. 283, NO. 29, pp. 20383–20396, : July 18, 2008 : 2)J Clin Invest. 2010;120(12):4289–4302. doi:10.1172/JCI42015.
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