人泡沫病毒GAG、ENV蛋白的原核表达、抗血清制备及鉴定-陕西师范大学学报期刊社网站

陕西师范大学学报（自然科学版）

化学与材料科学

人泡沫病毒GAG、ENV蛋白的原核表达、抗血清制备及鉴定

卢飞1，孙燕1，李晶2，李治1*

(1 陕西师范大学生命科学学院，陕西西安 710062；2 襄阳市中心医院感染性疾病科，湖北襄阳 441021)

卢飞，男，硕士研究生，主要研究方向为分子微生物.Email：feilu121@163.com.* 通信作者：李治，男，副教授，博士. Email：mmbiolab@gmail.com.

摘要:

构建了HFV的GAG和ENV蛋白原核表达载体pET32agag和pET32aenv，转化E.coli BL21 Star (DE3)后用IPTG诱导出高水平的GAG、ENV融合蛋白表达.以融合蛋白免疫新西兰兔制备了GAG和ENV抗血清.Western Blot检测表明，抗血清可以识别原核表达的GAG和ENV蛋白，说明抗血清具有较好的特异性.结合Western Blot和间接免疫荧光实验，表明抗血清可以检测到病毒表达的GAG和ENV蛋白.

关键词：

人泡沫病毒； GAG蛋白； ENV蛋白；原核表达；抗血清

收稿日期：

2011-05-17

中图分类号：

R392.11

文献标识码：

文章编号：

16724291(2012)03007105

基金项目：

教育部科学技术研究重点项目（108184）；国家自然科学基金资助项目（31000086）.

Doi:

Prokaryotic expression, purification, and antiserum preparation of GAG and ENV proteins of human foamy virus

LU Fei1, SUN Yan1, LI Jing2, LI Zhi1*

(1 College of Life Science, Shaanxi Normal University, Xi′an 710062, Shaanxi, China; 2 Department of Infection Disease, Xiangyang Central Hospital, Xiangyang 441021, Hubei, China)

Abstract:

In order to detect the efficiency of human foamy virus infection on protein expression level, the recombinant prokaryotic expression vectors pET32aGAG and pET32aENV were constructed successfully and transformed into E.coli BL21 Star (DE3), and the GAG, ENV fusion proteins were induced by IPTG and expressed at high level. Subsequently, the New Zealand rabbits were immunized with the fusion proteins for preparing the antiserums. Western Blot analysis showed that the antiserum could interact with GAG and ENV fusion proteins specifically. Based on the results from Western Blot and indirect immunofluotesent method, it is concluded that the antiserums were verified to interact with the natural GAG and ENV expressed from the viruses.

KeyWords:

human foamy virus; GAG protein; ENV protein; prokaryotic expression; antiserum