传染性造血组织坏死病毒核衣壳蛋白的原核表达及抗原性分析

应用RT-PCR方法扩增了长度为1176 bp的IHNV-ZYX株编码核衣壳(N)蛋白基因,将N基因克隆至原核表达载体pET30b,并在大肠杆菌Rosetta(DE3)中得到了表达.通过SDS-PAGE分析表明,重组菌诱导后得到了预期大小约48 KD的N蛋白,与理论值相符；提取N蛋白的包涵体,并制备抗血清.间接ELIS...

Prokaryotic expression and antigenicity analysis of the nucleocapsid gene of infectious hematopoietic necrosis virus

The gene encoding the viral nucleocapsid(N) was amplified by RT-PCR from IHNV-ZYX strain and cloned into pET30b vector.The expression of recombinant plasmid pET30b-N in E.coli Rosetta(DE3) was induced and detected by SDS-PAGE analysis.The molecular weight of expressed recombinant N protein was approximately 48 KD as predicted.The inclusion body containing fusion protein was extracted and immunized in rabbits to obtain the antisera against N protein.Antigenicity of N protein was analyzed by indirect ELISA and Western-blotting.The results showed that the expressed N protein has immunogenical and antigenical characters as native IHNV N protein.This study has laid an important material foundation for further studying the N gene function of IHNV-ZYX in immune protection,establishing sensitive and efficient methods in detecting infectious hematopoietic necrosis,and manufacturing genetic engineering vaccine.