论文部分内容阅读
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。
Objective: To construct IL-2 / pUC19 recombinant plasmid. Methods: mRNA was extracted from PHA-stimulated human Jurkat cells and IL-2 gene was obtained by RT-PCR. The cloned plasmids pUC19 and IL-2 were digested with BamHⅠand EcoR Ⅰ and purified by low melting point gel. The product was transformed into DH5a competent bacteria, screened colonies and identified by sequencing. Results: The expected IL-2 band was obtained by electrophoresis and sequencing confirmed the correct IL-2 sequence. Conclusion: The IL-2 / pUC19 recombinant plasmid can be constructed easily and efficiently by using the cloning plasmid pUC19.