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[目的]提高gG1基因中植物的表达量,研制Ⅰ型单纯疱疹病毒的口服植物疫苗。[方法]PCR扩增植物组成型表达启动子CaMV35S、378 bp的单纯疱疹病毒gG1基因、NOS终止子基因,将这些基因插入植物表达载体pCAMBIA1301的多克隆位点处,将pCAMBIA1301-gG1转化至大肠杆菌DH5α,用双酶切、测序进行鉴定。最后,将其转化至农杆菌EHA105,用PCR进行鉴定。[结果]从转化农杆菌中提取pCAMBIA1301-gG1DNA中得到了预期大小的gG1基因。[结论]成功构建了单纯疱疹病毒gG1基因的植物表达载体,为转化植物奠定了基础。
[Objective] To improve the expression of plant in gG1 gene and develop oral plant vaccine of herpes simplex virus type 1. [Method] The plant constitutive expression promoter CaMV35S and 378 bp of herpes simplex virus gG1 gene and NOS terminator gene were amplified by PCR. These genes were inserted into the multiple cloning site of plant expression vector pCAMBIA1301 and transformed into pCAMBIA1301-gG1 Bacillus DH5α, double digestion, sequencing identification. Finally, it was transformed into Agrobacterium EHA105 and identified by PCR. [Result] The gG1 gene of the expected size was obtained from pCAMBIA1301-gG1 DNA extracted from Agrobacterium tumefaciens. [Conclusion] The plant expression vector of herpes simplex virus gG1 gene was successfully constructed, which laid the foundation for plant transformation.