摘要
目的 构建含狂犬病毒 3a G株糖蛋白的重组质粒p CI- neorab G,为进一步表达及 DNA免疫做准备 .方法 从感染了狂犬病毒 3a G株的鼠脑组织中提取总 RNA进行 RT-PCR,扩增出编码糖蛋白的基因片段 ,重组入 p UC19克隆载体 .再将 p UC19rab G中的糖蛋白基因片段亚克隆入 p CI- neo真核表达载体 .结果 从 3a G株基因组中扩增出特异的糖蛋白基因片段 ,克隆成功 p UC19rab G;经亚克隆 ,筛选鉴定获得了 p CI- neorab G 重组质粒 .结论 构建成功狂犬病毒p U C19rab G重组质粒 ,亚克隆成功 p CI- neorab G重组质粒 ,为下一步表达及
AIM To prepare expression and DNA immunization by constructing recombinomt plasmid pCI neorabG containing glycoprotein gene of rabies virus 3aG strain. METHODS Total RNA were extracted from rabies virus 3aG infected mouse cerebral tissues, then glycoprotein gene fragment was amplified by RT-PCR and cloned into pUC19 plasmid (named pUC19rabG). The glycoprotein gene was then subcloned into eukaryotic expression vector pCI-neo. RESULTS Specific glycoprotein gene of rabies virus 3aG strain was successfully amplified and cloned into pUC19rabG. Fur-thermore, a recombinant plasmid pCI-neorabG for eukaryotic expression was also obtained after subcloning, selection and confirmation. CONCLUSION The obtaining of pUC19rabG and pCI-neorabG plasmids has laid a solid foundation for expression and DNA immunization.
出处
《第四军医大学学报》
2000年第7期900-902,共3页
Journal of the Fourth Military Medical University
