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TA Cloning® Kit, with pCR™2.1 Vector and One Shot® TOP10F' Chemically Competent E. coli, no manual - FAQs

查看更多产品信息 TA Cloning® Kit, with pCR™2.1 Vector and One Shot® TOP10F' Chemically Competent E. coli, no manual - FAQs (KNM203040)

2 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.