动物医学进展2016，Vol.37Issue(10)：34-40,7.
猪瘟病毒C株特异的TCRVα5和Vβ6真核表达载体的构建及共转染研究
Construction and Cotransfection of Eukaryotic Expression Plasmids TCR Vα5 and Vβ6 Specific for CSFV-C Strain
摘要
Abstract
Previously,we detected TCR Vα5 and TCR Vβ6 gene families specific for CSFV-C strain.In this study,to further observe the expression level in vitro ,RT-PCR technique was used to amplify the full-length sequences of TCR Vα5 and TCR Vβ6,and recombinant plasmids of TCR Vα5-pIRES2-EGFP and TCR Vβ6-pIRES2-EGFP were also constructed and co-transfected into 293T cells to observe the expression of CSFV-C specificαβTCR gene.In result,the TCR Vα5 and TCR Vβ6 sequences were 816 bp and 945 bp, respectively;the sequences of TCR Vα5-pIRES2-EGFP and TCR Vβ6-pIERS2-EGFP recombinant plasmids were confirmed to be correct by restriction enzyme digestion analysis and sequencing.Green fluorescence was observed with at least 70% transfected cells after 24 h post-transfection.Besides,the expression of TCR Vα5 and TCR Vβ6 were detected through real-time PCR and the EGFP protein was also verified by Western blot and the EGFP expression level was the same between two groups.These results indicated that the eukaryotic expression vector of TCR Vα5-pIRES2-EGFP and TCR Vβ6-pIRES2-EGFP specific for CSFV-C strain were constructed successfully and can be co-transfected into the 293T cells,realizing co-ex-pression of TCR Vα5 and TCR Vβ6 gene in vitro .关键词
猪瘟病毒C株/TCR Vα/Vβ基因/脂质体转染/增强绿色荧光蛋白Key words
Classical swine fever-C strain/TCR Vα/Vβgene/lipofectin transfection/EGFP分类
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王春燕,陈国华,何小兵,曾爽,贾怀杰,房永祥,冯园,李守杰,景志忠..猪瘟病毒C株特异的TCRVα5和Vβ6真核表达载体的构建及共转染研究[J].动物医学进展,2016,37(10):34-40,7.基金项目
国家自然科学基金项目 （）
