羊口疮病毒B2L基因克隆及表达

摘要

为了获得羊口疮病毒囊膜蛋白。以羊口疮病毒基因组为模板，利用羊口疮病毒B2L基因特异性引物，进行PCR扩增，回收以及纯化PCR产物，连接到PMD18-T载体上，测序结果表明B2L基因全长1137 bp，编码379个氨基酸。再将B2L基因克隆至pET-30a，构建原核表达载体pET30a-B2L。经菌液PCR、酶切鉴定以及SDS-PAGE等方法表明成功构建原核表达载体，这将为今后的单抗制备以及羊口疮病毒的疫苗研究打下基础。%To obtain ORFV envelope protein and take ORFV genome as template,ORFV B2L gene was used as specific primers to amplify PCR and recovery and purify PCR products. Connected PCR products to PMD18-T vector,the sequence result showed that full-length of B2L gene was 1137 bp,and Encoding was 379 amino acids. B2L gene was cloned into pET-30a to construct prokaryotic expression vector pET30a-B2L. Prokaryotic expression vector was constructed by the methods of bacterium liquid PCR, enzyme identification and SDS-PAGE,which would provide the research basis of monoclonal antibody and ORFV vaccine in the future.