摘要
将从组织病料中提取的鸡传染性贫血病毒(chicken infectious anemia virus,CIAV)核酸进行PCR扩增,获得vp1基因,将其克隆到表达载体pET32a(+)中,构建了CIAVvp1基因重组质粒,命名为pET32a-vp1。将pET32a-vp1转化E.coliplys,重组菌经IPTG诱导,SDS-PAGE分析结果表明,vp1基因在大肠杆菌中成功表达。纯化的蛋白质作为包被抗原,ELISA鉴定结果表明具有良好的抗原性。本试验结果为进一步研究用重组抗原制备CIAV ELISA诊断试剂盒奠定了基础。
We extracted chicken infectious anemia virus （CIAV） genome from liver tissue, amplified and gained vpl gene using the polymerase chain reaction （PCR）. By using the recombinant DNA technology, we digested insert DNA and pET-32a （＋） with restriction enzyme BamH I and Xho I , and then purified pET32a（＋） vector DNA and insert DNA, after that liga- ting pET32a（＋） vector DNA and insert DNA. Transform recombinant pET32a-vpl into non expression host E. coli DH5a competent cells. Using colony PCR to identify positive clones and verify reading frame by sequencing. Transform the plasmid pET32a-vpl into host E. coli plys. The recombinant bacteria was induced by IPTG and analyzed with SDS-PAGE. The results showed that vpl gene was expressed in E. coli at high level. ELISA identified target protein was vpl fusion protein. The pres ent study would be helpful for the development of ELISA diagnostic kit with recombinant antigen of CIAV.
出处
《中国畜牧兽医》
CAS
北大核心
2013年第8期34-38,共5页
China Animal Husbandry & Veterinary Medicine
基金
太原市科技项目资助(12-0149)
关键词
鸡传染性贫血病毒
VP1
原核表达
chicken infectious anemia virus（CIAV）
vpl
prokaryotic expression
