单纯疱疹病毒gG1基因植物表达载体的构建

Construction of plant expression vector of gG1 gene of Herpes simplex virus

[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 the gGl gene in peanuts and develop oral vaccine of Herpes simplex virus type I （HSV -1 ）. [ Methods]The constitutive promoter CaMV35S gene,the gG1 gene and the terminator NOS gene were am- plified by Polymerase Chain Reaction （PCR） , afterwards, inserted into the multiple cloning site of pCAMBIA1301. The plant expression vector pCAMBIA1301 -gG1 was transformed into Escherichia coil,which was identified by double digestion and se- quences analysis. In the end,it was transformed into Agrobacterium EHA105, which was identified by PCR. [ Results] The ex- pected bands of gG1 gene from recombinant plasmid pCAMBIA1301 -gG1 in Agrobacterium EHA105 were obtained by agarose gel elec -trophoresis. [ Conclusion]The plant expression vector had been successfully constructed,which laid a solid founda- tion for the transformation of plants.