钠钾泵β2亚基C末端编码区酵母双杂交载体的构建与表达

Cloning and expression of Na^+,K^+-ATPase gene in yeast

以人胎儿脑cDNA文库为模板克隆了钠钾泵β2亚基C末端编码区(N KAβ2ct),构建了表达载体pGBKT 7-NKAβ2ct,并对其进行了表达,对表达产物进行了检测。结果表明,N KAβ2ct长690 bp,pGBKT 7-NKAβ2ct酵母表达载体构建成功,在酵母中表达良好,可用于后续的酵母双杂交文库筛选工作。

The C terminal of coding region of Na^＋ ,K^＋-ATPase beta2 subunit （NKAβ2ct） was cloned, and a recombined plasmid allowing the expression in yeast for further study of the function was constructed. The coding region of NKAβ2ct was amplified at first from human brain cDNA library by using PCR technique and inserted into the yeast expression vector pGBKT7 followed with Western blotting technique. Results showed that NKAβ2ct yeast expression was successfully cloned for the two-hybrid system analysis,which would advance the functional study of NKAβ2.