鸡传染性喉气管炎病毒gB基因的克隆及其原核表达

Cloning the gB Gene of ILTV Vaccine Strain and Its Prokaryotic Expression

根据鸡传染性喉气管炎病毒(ILTV)的gB基因序列和表达载体pBV221的多克隆位点,设计了1对引物。以ILTV疫苗株基因组为模板,应用PCR扩增出gB基因片段,然后将其定向克隆于原核表达载体pBV221中,构建了重组质粒pBV-gB。重组质粒转化到受体菌DH5α中,得到阳性重组菌株DH5α(pBV-gB)。该菌株在温控诱导下,SDS-PAGE电泳可见表达的特异蛋白。ELISA检测证明,表达的蛋白具有较好的反应原性。

gB gene was amplified by polymerase chain reaction(PCR) from ILTV vaccine strain′s genomic DNA.Sequence analysis indicated that ILTV gB gene was contained 2 622 bp.Then gB gene was inserted into pBV_ 221 vector via the multiple cloning sites EcoRⅠ/BamHⅠ.The analysis results with SDS-PAGE showed that gB glycoprotein was expressed in E.coli DH5α and recombinant glycoprotein expressed as inclusion body.The expression of gB gene provides the basis for preparation of vaccine and preventing chicks from ILTV infection.