TCR Vβ8/pcDNA3.1重组质粒的构建  

Construction of recombinant plasmid of TCR β8/pcDNA3. 1

作　　者：张明智[1] 赵国强[2] 李继昌[3] 董子明[4] 李学民[5] 

机构地区：[1]郑州大学第一附属医院肿瘤科,郑州450052 [2]郑州大学基础医学院免疫学教研室,郑州450052 [3]郑州大学第一附属医院消化内科,郑州450052 [4]郑州大学基础医学院病理生理学教研室,郑州450052 [5]郑州大学第一附属医院检验科,郑州450052

出　　处：《郑州大学学报（医学版）》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

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.

关 键 词：淋巴瘤 TCR 独特型 重组质粒 DNA疫苗 

分 类 号：R318.5[医药卫生—生物医学工程] R733.4[医药卫生—基础医学]