禽脑脊髓炎病毒内蒙分离株外壳蛋白VP1基因在大肠杆菌中的融合表达及活性分析

为研制禽脑脊髓炎病毒(avian encephalomyelitis virus,AEV)亚单位疫苗,对AEV-NH937内蒙分离株外壳蛋白VP1基因进行融合表达、纯化及活性分析.根据AEV-NH937基因组全序列,设计一对引物.利用RT-PCR技术,扩增AEV的外壳蛋白VP1基因.经序列分析,VP1基因编码区为810bp,编码270个氨基酸残基.将VP1基因插入载体pGEX-4T1构建表达重组质粒,然后转化大肠杆菌BL21.经IPTG诱导后,用聚丙稀酰氨凝胶电泳分析,结果表明,有融合蛋白表达,其分子量为56KD.纯化后,利用ELISA检测分析VP1蛋白,证明VP1蛋白有一定的抗原活性.

FUSION EXPRESSION AND ACTIVITY ASSAY OF VP1 CAPSID PROTEIN GENE OF AVIAN ENCEPHALOMYCLITIS VIRUS NH937 STRAIN ISOLATED IN INNIER MONGOLIA

The capsid protein VP1 gene of avian encephalomyclitis virus NH937 strain isolated in Inner Mongolia is expressed in E.coli BL21 in order to develop novel subunit vaccine.A pair of primers is designed according to AEV-NH937 strain.The capsid protein of VP1 gene is amplified by use of RT-PCR.Sequence analysis shows that the VP1 gene consists of 810bp,which encodes 270 amino acid residues.The expression recombinat plasmid is constructed by inserting VP1 gene into vector pGEX-4T1,and then transformed into E.coli BL21.SDS-PAGE analysis shows that the VP1 fusion protein is expressed as inclusion after induction with IPTG.The molecule of the VP1 fusion protein is 53 kD.After purification,with the help of ELISA analysis,the antigenicity of the VP1 protein has been detected.