摘要
用反转录聚合酶链式反应 (RT_PCR)技术 ,从轮状病毒感染的细胞中扩增 816bp、916bp的VP4、VP7片段中抗原表位区。通过T4DNA连接酶将其直接连接于克隆载体质粒pGEM_T上 ,转化至受体菌DH5α中。提取质粒经PCR扩增、酶切鉴定 ,证明重组质粒pT_V4、PT_V7中含有轮状病毒的VP4、VP7基因片段。经核苷酸序列分析 ,表明克隆了轮状病毒主要保护性抗原VP4。
The antigenic determinants of Rotavirus VP4 and VP7 genes were amplified from cell infected with rotavirus by reverse transcription_polymerase chain reaction(RT_PCR),the length of target genes was 81bp and 91bp.The products of RT_PCR were ligated with plasmid pGEM_T and transformated to E.coli DH5α.By the analysis of restriction endonuclease and PCR,the fragments of VP4 and VP7 were cloned and the recombinant plasmids PT_V4 and PT_V7 were constructed.The results of nucleotide sequencing showed that the inserted genes had positive reading frame.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2003年第2期99-103,共5页
Chinese Journal of Preventive Veterinary Medicine
