联系我们 加入收藏
 
年期检索 高级检索
33
2025年5月5日 星期一
  研究论文 本期目录 | 过刊浏览 | 高级检索 |
鸡传染性支气管炎病毒S1基因在大肠杆菌中的表达及其生物活性的初步鉴定
谢红梅 朱来华 常维山
山东农业大学动物科技学院 青岛出入境检验检疫局 山东农业大学动物科技学院
The expression of S1 gene of IBV in E.coli and the original identification of it’s biological activity
全文: PDF (469 KB)   HTML (1 KB) 
输出: BibTeX | EndNote (RIS)      
摘要 摘要:利用基因重组技术将鸡传染性支气管炎病毒S1基因分段(A段和B段)克隆到pGEX-6p-1载体中,并利用IPTG诱导表达。表达产物裂解后,经SDS-PAGE电泳和考马司亮蓝染色,有明显的融合表达蛋白条带。通过特异性抗体的Western-blot检测,两段表达产物均在相应位置出现了明显的条带,证明表达产物具有部分抗原性。用Glutathione Sepharose 4B纯化出GST融合蛋白。将纯化得到的融合蛋白免疫家兔,连续免疫3次,将得到的抗体与200TOCID50/0.1ml病毒工作液在37℃温育1h进行气管环试验,计算结果 A段抗体的50%血清中和终点为1:18,而B段抗体的50%血清中和终点为1:53,从而表明这两段的表达产物具有较好的免疫原性,且B段表达产物的免疫原性比A段高。
服务
把本文推荐给朋友
加入我的书架
加入引用管理器
E-mail Alert
RSS
作者相关文章
谢红梅朱来华常维山
关键词 大白菜甘蓝杂种一代小孢子培养异附加系    
Abstract:Abstract: The S1 gene of IBV was cloned into expression vector pGEX-6p-1 by gene recombination technology. E. coli transformed by the recombinant plasmid containing IBV S1gene was induced by IPTG. after the induced A E. coli were lysed, fusion proteins in pGEX-IBV-S1A and pGEX-IBV-S1B were revealed by SDS-PAGE. And, obvious bands in pGEX-IBV-S1A and pGEX-IBV-S1B were showed by Western-blot. The result demonstrated partial antigenicity of the expression products. GST fusion proteins of A and B were purified using Glutathione Sepharose 4B. Rabbits were vaccinated with the purified GST fusion proteins. The antibody and virus were incubated in 37℃ incubator for 30 minutes, and then TOC experiment was made. The result showed that the antibody induced by segment A could inhibit the partial activity of IBV with SN endpoint of 1:18 while the antibody induced by segment B could completely inhibit the activity of virus with SN endpoint of 1:53. It is proved that the immunogenicity of segment B was better than that of segments A.
Key wordsChinese cabbage    Cabbage    Hybrid    Microspore culture    alien addition lines
收稿日期: 2006-03-20     
通讯作者: 常维山   
引用本文:   
谢红梅 朱来华 常维山 . 鸡传染性支气管炎病毒S1基因在大肠杆菌中的表达及其生物活性的初步鉴定[J]. , 2007, 15(1): 107-111.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/      或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2007/V15/I1/107
 
版权所有 © 2014 《农业生物技术学报》编辑部   京ICP备11035905号-3
地址:北京市海淀区圆明园西路2号中国农业大学生命科学楼1053室 邮编:100193
电话:010-62733684 传真:010-62731615 E-mail: nsjxb@cau.edu.cn