| 鹅副粘病毒M基因主要抗原位点片段的原核表达及鉴定 |
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| 引用本文： | 张萍,韩秀娥,王占锋,魏萍.鹅副粘病毒M基因主要抗原位点片段的原核表达及鉴定[J].东北农业大学学报,2012(9):56-59. |
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| 作者姓名： | 张萍 韩秀娥 王占锋 魏萍 |
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| 作者单位： | 1. 东北农业大学动物医学学院,哈尔滨 150030
2. 东北农业大学国乳中心,哈尔滨 150030 |
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| 基金项目： | 东北农业大学创新团队项目(CXT00641) |
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| 摘 要： | 根据GenBank中发表的GPMV SF02株M基因核苷酸序列设计合成一对引物,通过RT-PCR扩增JS/1/97/G0株M基因主要抗原位点片段M1,将其克隆入pMD18-T载体,序列测定和分析表明,所扩增的M1基因核苷酸长为444 bp,共编码148个氨基酸.将M1同载体PGEX-6P-1连接后,转化入E.coli BL21(DE3)PlysS,经IPTG诱导表达出目标蛋白,与预计相符.对表达的M1蛋白进行Western blot鉴定,表明所表达的M1蛋白可以与GPMV SF02株阳性血清发生特异性反应.
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| 关 键 词： | 鹅副粘病毒M基因 原核表达 抗原性检测 |
Prokaryotic expression of major antigen site in M gene from goose paramyxovirus and detection |
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| ZHANG Ping,HAN Xiue,WANG Zhanfeng,WEI Ping.Prokaryotic expression of major antigen site in M gene from goose paramyxovirus and detection[J].Journal of Northeast Agricultural University,2012(9):56-59. |
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| Authors: | ZHANG Ping HAN Xiue WANG Zhanfeng WEI Ping |
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| Institution: | 1(1.School of Veterinary Medicine,Northeast Agricultural University,Harbin 150030,China;2.Dairy Industry Technical Development Center,Northease Agricultural University,Harbin 150030,China) |
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| Abstract: | In this study,according to M gene sequence of GPMV SF02 isolate strain published in GenBank,two pairs of primers were desighed and M1 gene was amplified by RT-PCR.The specific DNA products were cloned into pMD18-T vector.The sequence analysis showed that M1 gene was 444 bp in length,encoding 148 amino acid residues;the major antigen site of M gene was inserted into expression plasmid pGEX-6P-1.The recombinant plasmid pGEX-6P-M1 was transformed into E.coli BL21(DE3)PlysS and induced with IPTG.A fusion protein as we expected had been found.The expressed products were tested by western blot.The results showed that GST-M1 fusion protein had a positive reaction. |
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| Keywords: | goose paramyxovirus prokaryotic expression detection of antigenicity |
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