| 猪瘟病毒p80基因丝氨酸蛋白酶功能区的原核表达及小鼠免疫试验 |
| |
| 引用本文： | 鲁絮,许信刚,张彦明,张淼涛,李少方.猪瘟病毒p80基因丝氨酸蛋白酶功能区的原核表达及小鼠免疫试验[J].西北农业学报,2007,16(6):14-18. |
| |
| 作者姓名： | 鲁絮 许信刚 张彦明 张淼涛 李少方 |
| |
| 作者单位： | 1. 中国农业科学院兰州兽医研究所,农业部畜禽病毒学重点开放实验室,兰州,730046;西北农林科技大学动物科技学院,陕西杨凌,712100
2. 西北农林科技大学动物科技学院,陕西杨凌,712100 |
| |
| 基金项目： | 农业部重点实验室基金;西北农林科技大学博士科研启动经费 |
| |
| 摘 要： | 根据已发表的猪瘟病毒Alfort株的全基因组序列,设计2对引物以Shimen细胞毒株为材料,一步法提取总RNA,利用RT-PCR和套式PCR,成功扩增出p80全长基因,包括丝氨酸蛋白酶以及NTPase/RNA解旋酶功能区,大小约为2.0kb。将p80基因中的丝氨酸蛋白酶基因克隆到原核表达载体pET-32a中,获得重组质粒pET-p80。将重组质粒转化大肠杆菌Rosetta（DE3）中,IPTG诱导表达得到分子量约为43ku的目的蛋白,与理论大小相符。将菌体蛋白经Ni柱纯化,获得纯化重组p80蛋白,Western blot检测表明,表达的目的蛋白能与CSFV阳性血清发生反应。用纯化的p80蛋白免疫Bal b/c小鼠,成功制备了抗p80蛋白的抗血清。
|
| 关 键 词： | 猪瘟病毒 p80基因 原核表达 免疫 |
| 文章编号： | 1004-1389（2007）06-0014-05 |
| 收稿时间： | 2007-04-13 |
| 修稿时间： | 2007-06-15 |
Expression of Serine Proteinase Functional Domain of p80 Gene of CSFV in E. coli and Mice Immune Test |
| |
| LU Xu,XU Xin-gang,ZHANG Yan-ming,ZHANG Miao-tao and LI Shao-fang.Expression of Serine Proteinase Functional Domain of p80 Gene of CSFV in E. coli and Mice Immune Test[J].Acta Agriculturae Boreali-occidentalis Sinica,2007,16(6):14-18. |
| |
| Authors: | LU Xu XU Xin-gang ZHANG Yan-ming ZHANG Miao-tao and LI Shao-fang |
| |
| Institution: | State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China;College of Animal Science and Technology, Northwest A & F University, Yangling Shaanxi 712100, China;College of Animal Science and Technology, Northwest A & F University, Yangling Shaanxi 712100, China;College of Animal Science and Technology, Northwest A & F University, Yangling Shaanxi 712100, China;College of Animal Science and Technology, Northwest A & F University, Yangling Shaanxi 712100, China;College of Animal Science and Technology, Northwest A & F University, Yangling Shaanxi 712100, China |
| |
| Abstract: | Two pairs of primers were designed based on the sequence of CSFV Alfort strain.Serine protease and NTPase/ RNA helicase functional domain,about 2.0kb of p80 genes of CSFV Shimen strain were amplified by RT-PCR and nPCR.Serine protease gene was cloned into prokaryotic expression vector pET-32a.The inserted position,the orientation and the ORF of the recombinant plasmids pET-p80 were proven to be correct by PCR,restriction enzyme digestion and sequence analysis.The about 42ku target protein was expressed in E.coli Rosetta(DE3) induced with IPTG.Western-blotting showed that the expressed proteins could be recognized by CSFV positive serum.Balb/c mice were injected in abdomen with purified p80 protein and its antiserum was made successfully. |
| |
| Keywords: | CSFV Shimen strain p80 gene Prokaryotic expression Immune |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
| 点击此处可从《西北农业学报》浏览原始摘要信息 |
| 点击此处可从《西北农业学报》下载免费的PDF全文 |
|
