Search

  • 联系我们
  • 货号直购

TA Cloning™ Kit, Dual Promoter, with pCR™II Vector and One Shot™ TOP10F' Chemically Competent E. coli, 20 reactions - FAQs

查看更多产品信息 TA Cloning™ Kit, Dual Promoter, with pCR™II Vector and One Shot™ TOP10F' Chemically Competent E. coli - FAQs (K206001, K206040)

3 个常见问题解答

我可以使用TOP10F’感受态细胞对包含ccdB基因的TOPO载体进行转化吗？

含F质粒的菌株，如TOP10F’，不推荐用于转化和筛选任何含ccdB基因的TOPO载体的重组克隆。F质粒编码CcdA蛋白，该蛋白是CcdB旋转酶-毒素蛋白的抑制剂，但是CcdA蛋白的半衰期比CcdB蛋白短。当没有足够的CcdA抑制CcdB蛋白时，过表达CcdB蛋白会引起细胞死亡。

Can I use TOP10F' competent cells for transformation of my TOPO vector that contains the ccdB gene?

Strains that contain an F plasmid, such as TOP10F', are not recommended for transformation and selection of recombinant clones in any TOPO vector containing the ccdB gene. While the F plasmid does encode the CcdA protein, which acts as an inhibitor of the CcdB gyrase-toxin protein, the half-life of the CcdA protein is shorter than that of the CcdB protein. Overexpression of the CcdB protein causes cell death when its action is not prevented by sufficient CcdA.

Have you compared in vitro transcription levels between SP6 and T7 promoters in your pCRII vectors?

No, we have not done in-house comparisons of transcription levels. It is widely known though that T7 polymerase produces more RNA than SP6 (on the order of 10-fold higher).