| 人泡沫病毒GAG、ENV蛋白的原核表达、抗血清制备及鉴定 |
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| 引用本文： | 卢飞,孙燕,李晶,李治.人泡沫病毒GAG、ENV蛋白的原核表达、抗血清制备及鉴定[J].陕西师范大学学报,2012(3):71-75. |
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| 作者姓名： | 卢飞 孙燕 李晶 李治 |
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| 作者单位： | 陕西师范大学生命科学学院;襄阳市中心医院感染性疾病科 |
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| 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) |
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| 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 BL21Star(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.
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| 关 键 词： | 人泡沫病毒 GAG蛋白 ENV蛋白 原核表达 抗血清 |
Prokaryotic expression,purification,and antiserum preparation of GAG and ENV proteins of human foamy virus |
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| LU Fei,SUN Yan,LI Jing,LI Zhi.Prokaryotic expression,purification,and antiserum preparation of GAG and ENV proteins of human foamy virus[J].Journal of Shaanxi Normal University: Nat Sci Ed,2012(3):71-75. |
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| Authors: | LU Fei SUN Yan LI Jing LI Zhi |
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| Institution: | 1*(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) |
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| 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. |
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| Keywords: | human foamy virus GAG protein ENV protein prokaryotic expression antiserum |
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