摘要
目的:为获得抗SARS的转基因植物疫苗,进行了SARS S1基因的克隆,植物表达载体构建的研究。方法:根据SARS S蛋白受体结合结构域318-510氨基酸区域,设计合成长579 bp的序列片断(S1基因)。并以之为模板,进行PCR扩增,扩增产物被克隆至pGEM-Teasy载体进行序列测定。然后将S1基因插入植物表达载体pCAMB IA2301的35S启动子和NOS终止子之间,构建植物表达载体pCS1。将重组体转化大肠杆菌E.coliXL1,并对重组体进行了鉴定。结果:酶切鉴定和序列分析显示,克隆的目的基因与设计的片断序列一致;双酶切表明,植物表达载体的构建完全正确。结论:成功地克隆了S1基因和构建了含有S1基因的植物表达载体,为进一步获得抗SARS的转基因植物疫苗打下了基础。
Objective： To clone SARS coronavirus S1 gene and construct its plant expression vector and research and develop anti- SARS transgenic plant vaccine. Methods： A specific DNA fragment（S1 ） of 579 bp was synthesized and amplified by PCR according to SAILS spike protein receptor binding domain （318 -510AA） The PCR product was cloned into pGEM -Teasy vector and sequenced. The plant expression vector pCSlwas construtted by inserting S1 gene into between 35S promoter and NOS terminator. The recombinant DNA was trans- formed into E. coli XL1 and identified. Results： Enzyme digestion and sequencing analysis indicated the base se- quence of cloned gene was identical with the designed DNA fragment; double - enzyme digestion demonstrated that the constructed S1 plant expression vector was correct. Conclusion： The SI gene is cloned and its plant expression vector is successfully constructed, which lays a basis for the research and development of anti - SARS transgenic plant vaccine.
出处
《海南医学院学报》
CAS
2008年第6期605-607,共3页
Journal of Hainan Medical University
基金
海南省教育厅基金项目(No:Hjkj200421)
关键词
冠状病毒属
基因
遗传载体
聚合酶链反应
Coronavirus
Gene
Genetic vector
Polymerase chain reaction（PCR）
