IL-2/pUC19重组质粒的构建

Construction of recombinant plasmid of IL-2/pUC-19

目的:构建IL－2/pUC19重组质粒。方法:从PHA刺激的人Jurkat细胞中提取mRNA,经RT－PCR法获取 IL-2基因;将克隆质粒 pUC19和 IL－2基因行 BamHⅠ 和 EcoR Ⅰ双酶切、低熔点胶纯化、连接酶切产物、转化DH5a感受态细菌、筛选菌落和测序鉴定。结果:电泳获得预期的IL－2条带,测序证实为正确的IL－2序列。结论:应用克隆质粒 pUC19可方便高效地构建IL－2/pUC19重组质粒。

Aim: To construct recombinant plasmid of IL-2/pUC19. Methods: The mRNA of IL-2 was extracted from human Jurkat cells activated with PHA ; IL-2 gene was obtained by using RT-PCR method; the cloned plasmid pUC-19 and IL-2 gene were cleaved with 2 restriction endonucleases Bam HI and EcoRI. The products were separated and purified in low melting temprature agarose gels and linked and transformed into bacteria DH5a; the cl-onies were screened and the recombinant was squenced for identifying. Result: The expected band of IL-2 was obtained with eletrophoresis and the squence of IL-2 was confirmed by squencing analysis. Conclusion: Clone plasmid pUC-19 is used conveniently and effectively for constructing IL-2/ pUC-19.