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abbr_bff0ccfd6a54ee569e9f191ff3dec2ee.pdf
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2 c$ l1 j' I1 @& \8 H' d5 K6 e) ZGeneration of TF-inducible ES cell lines8 p7 s- v. y) U- D# b+ I/ R: j) O
The exchange vector (pZhcSfi) was modified to express puromycin resistant gene and C-terminal His6-
+ t k! r: x- ?$ KFLAG tag protein. For selected genes, we identified corresponding cDNA clones containing a full ORF' l& s1 y' s/ @
from the in-house NIA mouse cDNA collection (Carter et al., 2003; Sharov et al., 2003) and NIH* j* c3 v7 y( ~3 A; p
Mammalian Gene Collection (Gerhard et al., 2004). Some ORF fragments were originally obtained by$ Q* K- T5 P8 v
reverse transcription-PCR using ES cell RNAs. These full ORFs were subcloned into modified exchange3 O& w2 P% U# F0 L" [+ P
vector and verified by DNA sequencing. ES[MC1R(20)] cells carrying ROSA-TET locus were cotransfected
" U/ R2 F1 X( Dwith exchange vector and pCAGGS-Cre and selected in presence of puromycin.% j$ R2 g* K4 {7 P
% O3 ^8 \+ I' a; H8 m6 u$ }; @( e' M, ?原文附上
) C$ N' H# j' F- R+ K因为生物基础太弱，又刚转到干细胞
% Y+ u( k! L3 `/ _ ^不太看懂这里面的表达谱的数据叫什么TF诱导
8 ]. i$ T1 O" U* o0 F请高手不吝指教，在线等4 Y% \8 m3 v4 E( W6 Z
感谢了$ K7 L5 U- }2 ?# N7 U: E
数据名称为“Timed induction of 50 transcription factors in ES cells reveals a common mechanism to initiate differentiations” |
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发表于 2010-4-20 13:03
发表于 2010-4-20 19:41
