摘要
采用RT-PCR方法,从口蹄疫病毒中扩增出长约600 bp的核苷酸片段.纯化后与载体pMD18-Tsimple连接,重组质粒经PCR鉴定及DNA测序,结果表明克隆的片段为口蹄疫病毒L基因.将质粒pMD18-L与表达载体pEGFP-N1分别用BamHⅠ+XhoLⅠ双酶切,将所获目的基因与带有酶切位点的载体连接,经酶切、PCR鉴定及DNA测序,结果表明重组表达质粒pEGFP-L构建成功.将转染的BHK-21细胞,经荧光鉴定和RT-PCR检测,证实L基因在BHK-21细胞中得到表达.
The L gene of foot-and-mouth disease virus was amplified by RT--PCR,the purified DNA fragment was inserted into pMD18-T vector, which contains complete L gene. The plasmid pMD18-L and the expression vector pEGFP-N1 was digested by BamH I-t-XhoL I respectively, and the amplicon was cloned into＇pEGFP-N1 to construct recombinant expression plasmid. The recombinant plasmid was checked by restriction enzyme, PCR analysis and nucleic transfected with pEGFP-L. The expression proteins sults showed that proteins of L was expressed in BH were acid sequencing. Further BHK-21 cells were analyzed by fluoroscopy and RT-PCR. The re- K-21 cells.
出处
《甘肃农业大学学报》
CAS
CSCD
2007年第5期1-4,共4页
Journal of Gansu Agricultural University
基金
国家重点基础研究发展计划(973)项目(2005CB523201)
