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截短的猪生殖与呼吸综合征病毒M基因的
原核表达
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戚亭，崔尚金*，张超范
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(中国农业科学院 哈尔滨兽医研究所 兽医生物技术国家重点实验室
猪传染病研究室，黑龙江 哈尔滨150001)
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摘要：
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根据GenBank上登录的猪生殖与呼吸综合征病毒(PRRSV)基因组序列，设计了1对引物用于扩增PRRSV M基因，经PCR、酶切及测序鉴定，成功构建了PRRSV M基因原核表达载体。重组菌经IPTG诱导表达，SDS-PAGE和Western-blotting结果显示,出现一分子质量约为27.3ku的目的蛋白带，该表达蛋白可与PRRSV猪阳性血清发生特异性反应。表明，表达的蛋白具有良好的特异性。 |
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关键词：
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猪生殖与呼吸综合征病毒；M蛋白；原核表达 |
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中图分类号：S 852.659.6 文献标识码：
A 文章编号：1673-4696(2008)06-0457-04 |
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Expression of truncated gene-encoding M protein of PRRSV
in Escherichia coli |
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QI Ting,CUI Shang-jin,ZHANG Chao-fan |
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(National Key Laboratory of Veterinary Biotechnology/Division of Swine Infectious Disease,
Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150001,China)
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Abstract: According to the genomic sequence of porcine reproductive and respiratory syndrome virus(PRRSV) available in GenBank,a pair of primers were designed to amplify the sequence of gene-encoding M protein of PRRSV.The prokaryotic expression vector for M protein was successfully identified by PCR,enzymatic digestion and sequencing.The recombinant bacteria were induced by IPTG.A protein band of 27.3ku in molecular mass was observed in SDS-PAGE analysis.The specific band could react with PRRSV positive pig serum in Western-blotting.The results showed that the recombinant protein had good specificity with
anti-PRRSV serum. |
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Key words: porcine reproductive and respiratory syndrome virus;M protein;prokaryotic expression |
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