摘要
目的:构建重组质粒TCR Vβ8/pcDNA3．1。方法;从人淋巴瘤 Jurkat细胞中提取 mRNA,经 RT－PCR法获取 TCR Vβ8基因;将表达质粒 pcDNA3．l和 TCR Vβ8基因行 BamHI和 HindⅢ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 523 bp的 TCR Vβ8预期条带,测序证实为正确的 TCR Vβ8目的基因序列。结论:测序是构建TCR Vβ8/pcDNA3．l重组质粒的重要步骤。
Aim: To construct recombinant plasmid of TCRV β8/pcDNA3. 1. Methods:The mRNA of TCRV β8 was extracted from human Jurkat cells; gene of TCRV β8 was obtained by using RT-PCR method; the expressing plasmid pcDNA3. 1 and TCRV β8 genes were cleaved with 2 restriction endonucleases BamH Ⅰ and Hind Ⅲ. The products were seperated and purified in low melting temprature agarose gels and linked with each other. The recombinant plasmids were transformed into bacteria DH5a; the clonies were screened and the recombinant was sequenced for identification. Result; The expected 523bp band of TCR Vβ8 was obtained by eletrophoresis . The squence of TCR Vβ8/pcDNA3. 1 was confirmed by spquencing analysis. Conclusion: Seqencing is key step of constructing the recombinant plasmid of TCR Vβ8/pcDNA3. 1.
出处
《郑州大学学报（医学版）》
CAS
北大核心
2002年第3期268-273,共6页
Journal of Zhengzhou University(Medical Sciences)
基金
博士基金资助课题
