| 鸡贫血病毒衣壳蛋白VP1基因的克隆、原核表达和纯化 |
| |
| 引用本文： | 吴艳红,陈建龙,李丛丛,何成强,李云龙.鸡贫血病毒衣壳蛋白VP1基因的克隆、原核表达和纯化[J].动物医学进展,2007,28(1):46-49. |
| |
| 作者姓名： | 吴艳红 陈建龙 李丛丛 何成强 李云龙 |
| |
| 作者单位： | 1. 山东师范大学生命科学学院动物抗性实验室,山东济南,250014
2. 山东省济南第一中学,山东济南,250011 |
| |
| 摘 要： | 根据GenBank中CAV哈尔滨分离株基因序列,设计出针对CAV VP1大片段(608 bp)的引物,利用PCR方法从CAV基因组序列中扩增出VP1基因的大片段(608 bp),按照正确的读码框克隆到原核表达载体pET32a( )上,得到含VP1大片段的pET32a( )重组子,转化大肠埃希菌BL21(DE3)感受态细胞,IPTG诱导重组蛋白表达,经SDS-PAGE和Western blotting检测发现有分子质量约42 ku的融合蛋白表达,与预期分子质量大小一致,通过Ni亲和层析柱纯化出融合蛋白,经Western blotting鉴定,融合蛋白与His单抗能够结合.CAV VP1基因的克隆表达及其融合蛋白的纯化为后续制备多克隆抗体和单克隆抗体提供了良好的抗原来源,也为研究VP1与其他CAV蛋白之间的相互关系奠定了基础.
|
| 关 键 词： | 鸡贫血病毒 VP1 原核表达 |
| 文章编号： | 1007-5038（2007）01-0046-04 |
| 收稿时间： | 2006-10-08 |
| 修稿时间： | 2006年10月8日 |
Cloning of CAV VP1 Gene and Prokaryotic Expression and Purification of CAV VP1 Envelope Protein |
| |
| WU Yan-hong,CHEN Jian-long,LI Cong-cong,HE Cheng-qiang,LI Yun-long.Cloning of CAV VP1 Gene and Prokaryotic Expression and Purification of CAV VP1 Envelope Protein[J].Progress In Veterinary Medicine,2007,28(1):46-49. |
| |
| Authors: | WU Yan-hong CHEN Jian-long LI Cong-cong HE Cheng-qiang LI Yun-long |
| |
| Abstract: | A pair of primers were designed and synthesized according to the large fragment of CAV gene se- quence,and a 608 bp of VP1 gene fragment was amplified by PCR method from CAV genome and cloned into pET32a( ) vector,an E.coli expression vector,to construct a recombinant plasmid pET32a large fragment of VP1.The plasmid was transformed into E.coli BL21 (DE3) to express VP1 protein with IPTG induction,and a novel protein with molecular weight 42 ku was detected by SDS-PAGE and Western-blot- ting.The protein VP1 fused with His tag was purificated by His Bind Resin column,then the protein VP1 fused with His tag was detected by Western-blotting.The expressed product showed good reactivity to an- ti-His tag antibody.Cloning of CAV VP1 gene and the expression and purification of envelope protein VP1 provide the source of antigen for preparing polyclonal VP1 antibody and monoclonal VP1 antibody,and lay foundatioan for strudying the relationship among three CAV protein. |
| |
| Keywords: | Chicken anemia virus VP1 prokaryotic expression |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
|
