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[目的]构建人α防御素6(HD-6)酵母表达载体,为研究和解析人α防御素6的功能准备了条件。[方法]采用PCR方法,设计引物从cDNA文库中扩增出人α防御素6基因片段,将产物分离纯化后经EcoRⅠ和XbaⅠ酶切,并将其插入到毕赤酵母表达载体pPICZαA中,以得到重组的酵母表达载体pPICZαA/HD-6,最后进行琼脂糖电泳和酶切鉴定。[结果]从cDNA文库中扩增出的HD-6基因片断大小正确;电泳和酶切结果均证明已将此片段克隆到酵母表达载体pPICZαA内。[结论]成功地构建了人α防御素6基因的酵母表达载体pPICZαA/HD-6。
[Objective] To construct human alpha-defensin 6 (HD-6) yeast expression vector and prepare the conditions for the study and analysis of the function of human alpha-defensin 6. [Method] The human α-defensin 6 gene was amplified from the cDNA library by PCR. The product was isolated and purified by EcoR Ⅰ and Xba Ⅰ digestion and inserted into the expression vector pPICZαA of Pichia pastoris The recombinant yeast expression vector pPICZαA / HD-6 was obtained, and finally agarose gel electrophoresis and enzyme digestion were carried out. [Result] The size of HD-6 gene amplified from cDNA library was correct. The results of electrophoresis and digestion proved that the fragment was cloned into yeast expression vector pPICZαA. [Conclusion] The yeast expression vector pPICZαA / HD-6 of human α-defensin 6 gene was successfully constructed.