MDR1真核表达载体的构建及鉴定

Construction and Identification of Eukaryotic Expression Vector MDR1

采用PCR技术扩增MDR1基因全长序列及胞外区约1kb片段,并定向克隆到pcDNA3质粒载体CMV启动子序列下游的BamHⅠ和XhoⅠ限制性内切酶酶切位点之间。重组体经转化E coliJM109后可大量扩增,提取该重组体进行限制性内切酶分析,得到预期的目的片段,并且以重组体为模板,用特异性引物可扩增出MDR1基因的1kb片段。

<Abstrcat> The MDR1 gene span sequence and 1kb segment of cell outskirt were enlarged by PCR technology and were cloned to pcDNA3 plasmids CMV sequence down between BamHⅠ and XhoⅠ restrictive inner cut enzyme location point. Recombinant plasmids were enlarged in bulk by E.coli JM109. The recombinant plasmids were distilled and cut by restrictive inner cut enzyme. Expectant aim segment was seen. The 1kb segment was enlarged by PCR with recombinant.