| 传染性造血组织坏死病毒糖蛋白基因的原核表达及抗原性分析 |
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| 引用本文： | 赵永欣,赵丽丽,刘巍巍,王健楠,李一经,乔薪瑗,葛俊伟,刘敏.传染性造血组织坏死病毒糖蛋白基因的原核表达及抗原性分析[J].东北农业大学学报,2011,42(12):81-85. |
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| 作者姓名： | 赵永欣 赵丽丽 刘巍巍 王健楠 李一经 乔薪瑗 葛俊伟 刘敏 |
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| 作者单位： | 1. 东北农业大学动物科学技术学院,哈尔滨,150030
2. 东北农业大学动物医学学院,哈尔滨,150030 |
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| 基金项目： | 黑龙江省自然科学基金，黑龙江省教育厅科技项目 |
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| 摘 要： | 应用RT-PCR方法扩增了传染性造血组织坏死病毒IHNV-ZYX株编码的糖蛋白(G)基因,将G基因克隆至原核表达载体pET30b,并在大肠杆菌Rosetta( DE3)中得到了表达.SDS-PAGE分析表明,重组菌诱导后得到了预期大小的G蛋白.提取G蛋白的包涵体,并制备抗血清.间接ELISA和Western-blott...
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| 关 键 词： | 传染性造血组织坏死病毒 G基因 克隆表达 抗血清 |
Prokaryotic expression and antigenicity analysis of the glycoprotein gene of infectious hematopoietic necrosis virus |
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| ZHAO Yongxin,ZHAO Lili,LlU Weiwei,WANG Jiannan,LI Yijing,QIAO Xinyuan,GE Junwe,LIU Min.Prokaryotic expression and antigenicity analysis of the glycoprotein gene of infectious hematopoietic necrosis virus[J].Journal of Northeast Agricultural University,2011,42(12):81-85. |
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| Authors: | ZHAO Yongxin ZHAO Lili LlU Weiwei WANG Jiannan LI Yijing QIAO Xinyuan GE Junwe LIU Min |
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| Institution: | 1(1.College of Animal Sciences and Technology,Northeast Agricultural University,Harbin 150030,China;2.College of Veterinary Medicines,Northeast Agricultural University,Harbin 150030,China) |
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| Abstract: | The gene encoding the viral glycoprotein(G) was amplified by RT-PCR from IHNV-ZYX strain and cloned into pET30b vector.The expression of recombinant plasmid pET30b-G in E.coli Rosetta(DE3) was induced and detected by SDS-PAGE analysis.The molecular weight of expressed recombinant G protein was approximately 52 ku as predicted.The inclusion body containing fusion protein was extracted and immunized in rabbits to obtain the antisera against G protein.Antigenicity of G protein was analyzed by indirect ELISA and Western-blotting.The results showed that the immunogenical and antigenical characters of the expressed G protein was in common with native IHNV G protein.This study laid an important material foundation for further studying on the G gene function of IHNV-ZYX in immune protection,establishing sensitive and efficient methods in detecting infectious hematopoietic necrosis,and manufacturing genetic engineering vaccine. |
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| Keywords: | infectious hematopoietic necrosis virus(IHNV) glycoprotein gene cloning and expression antisera |
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