CA Jia-Li, CA Bao-Xiang and JIANG Beng. Expression of the M and N Protein Gene of Porcine Reproductive and Respiratory Syndrome Virus in E .coli[J]. Virologica Sinica, 1999, 14(3): 244-248.
Citation:
CA Jia-Li, CA Bao-Xiang, JIANG Beng.
Expression of the M and N Protein Gene of Porcine Reproductive and Respiratory Syndrome Virus in E .coli .VIROLOGICA SINICA, 1999, 14(3)
: 244-248.
猪繁殖和呼吸综合征病毒膜蛋白和核衣壳蛋白基因在大肠杆菌中的表达
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1.
南京农业大学动物医学院,南京210095
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2.
西南农业大学动物养殖学院,重庆400716
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摘要
以PRRSV弱毒株膜蛋白(M)和核衣壳(N)蛋白基因为模板,设计的一对含有EcoRI和BamHI酶切位点的引物,通过RTPCR扩增出一约900bp的MN基因片段,将此基因片段成功克隆于高效表达载体pBV220,构建成重组质粒pBVMN,导入大肠杆菌DH5,经温敏诱导,成功地表达了MN基因。表达产物经SDSPAGE电泳和Westernblot印迹分析,其分子量约34kD,与兔抗PRRSV高免血清发生反应,经光密度扫描分析,表达产物量占菌体总蛋白的12%。该研究为PRRS基因诊断抗原的研制奠定了基础
Expression of the M and N Protein Gene of Porcine Reproductive and Respiratory Syndrome Virus in E .coli
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Abstract
The primers for RT PCR were designed on the basis of M and N gene sequence of PRRSV. A gene fragment about 900 bp, which has digestion sites of EcoRI and BamHI, was amplified by RT PCR. The M and N gene were cloned into expression vector pBV220 and one recombinant PBVMN was constructed which highly expressed a 34 kD protein in \%E. coli\% DH 5 . The expressed product was identified by SDS PAGE and Western blotting, which occupies 12% of total bacterial protein. The report has laid a basis for the development of molecular diagnostic antigen of PRRSV.
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References
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Proportional views
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