甜菜夜蛾多核壳型核型多角体病毒vp39基因的克隆与表达(英文)

Cloning and expression of the vp39 gene of Spodoptera exigua multiplenucleocapsid nuclear polyhedrosis virus

参照苜宿银纹夜蛾核型多角体病毒 ( Ac MNPV)衣壳蛋白基因 vp3 9的序列设计引物 ,采用PCR技术扩增了甜菜夜蛾核型多角体病毒的约 1 .3 kb片段。通过将该片段亚克隆至原核表达载体p ET-2 8构建出重组表达质粒 p ET-Se3 9.以 p ET-Se3 9转化 E.coli BL2 1 .经 IPTG诱导后 ,Se MNPVvp3 9基因高效表达 .SDS-PAGE分析显示表达产物的分子量为 3 9KD,并且表达量在 IPTG诱导 4 h达到最高水平 .

A fragment of SeMNPV (Spodoptera exigua multiplenucleocapsid nuclear polyhedrosis virus) about 1.3 kb had been amplified by employing PCR technique and by designing a pair of primers based on the sequence of capsid protein gene vp39 of AcMNPV( Autographa californica multiplenucleocapsid nuclear polyhedrosis virus). The recombinant expression plasmid pET Se39 has been constructed by subcloning the 1.3 kb fragment into the procaryotic expression vector pET 28. After transformed into E.coli BL21, SeMNPV vp39 gene was highly expressed by the inducement of IPTG. SDS PAGE analysis showed that the molecular weight of the expressed protein was about 39 kD.The amount of the expressed product reached to the highest level when it had been induced with IPTG for 4 hours.