|
|
|
猪传染性胃肠炎病毒S基因A抗原位点序列的
克隆与原核表达
|
|
|
|
张春叶1，张莉3，沈红1，李焕荣1，吴国娟1，于同泉2，路苹2
|
|
|
|
（1.北京农学院 动物科学技术系，北京102206；2.农业应用新技术北京市重点实验室，北京102206；
3.北京市农林科学院 畜牧兽医研究所，北京100097）
|
|
|
|
摘要：
|
|
将RT-PCR扩增的猪传染性胃肠炎病毒(TGEV)S基因A抗原位点目的片段克隆、双酶切后连接于表达载体，经酶切、PCR和测序鉴定的阳性重组质粒转化BL21(DE3)，用IPTG诱导,进行SDS-PAGE和Western-blotting分析，确定目的蛋白的原核表达情况和免疫特异性。结果显示，目的片段的大小为534bp，序列分析表明，该基因与其他TGEV相应基因具有很高的同源性；Western-blotting检测可见分子质量约43ku的融合蛋白条带，表明，该蛋白具有良好的反应原性。 |
|
关键词：
|
|
猪传染性胃肠炎病毒；S基因A抗原位点；克隆；原核表达 |
|
中图分类号：S 852.659.6 文献标识码：
A 文章编号：1673-4696(2007)10-0845-05 |
|
|
Cloning and prokaryotic expression of antigenic site A sequence of
S gene of swine transmissible gastroenteritis virus |
|
|
|
ZHANG Chun-ye1,ZHANG Li3,SHEN Hong1,LI
Huan-rong1,WU Guo-juan1,YU Tong-quan2,LU
Ping2 |
|
|
|
（1.Department of Animal Science and Technology,Beijing University of Agriculture,Beijing 102206,China;
2.Beijing Key Laboratory of New Applicable Technology to Agriculture,Beijing 102206,China;3.Institute of Animal
Science and Veterinary Medicine,Beijing Academy of Agricultural and Forestry Sciences,Beijing 100097,China） |
|
|
Abstract: Antigenic site A sequence of S gene of swine transmissible gastroenteritis virus(TGEV) was amplified by RT-PCR and cloned into the prokaryotic expression vector after digestion with restriction enzyme.The positive recombinants identified by enzymatic digestion,PCR and sequencing was transformed into Escherichia coli BL21 and the recombinant transformant was induced with IPTG.The prokaryotic expression and the immunologic specificity of the expressed protein were analyzed by SDS-PAGE and Western-blotting.The results showed that the target gene was 534bp in length and had a relatively high homology with the corresponding gene from other TGEV strains.The expressed fusion protein was 43ku in size,and had good reactogenicity as revealed by Western-blotting. |
|
|
|
Key words: swine transmissible gastroenteritis virus;antigenic site A of S gene;cloning;prokaryotic expression |
|
|
|
|
|
《中国兽医科学》 ©
版权所有
Chinese Veterinary Science
zgsykx@zgsykx.com |
