【摘要】 目的:为了提高目的基因在番茄果实中的表达量,为进一步研究口服植物疫苗打下基础。方法:PCR扩增1.1kb番茄果实特异性启动子E8基因、460bp风疹病毒抗原E1基因、256bp NOS基因,将这些片段插入到pCAMBIA1301多克隆位点,得到植物表达载体pCAM1301E8-E1,经测序鉴定正确,将其转化至根癌农杆菌EHA105,然后进行酶切鉴定。结果:重组质粒酶切鉴定均得到预期片段,测序结果正确。结论:该实验成功构建番茄果实特异性启动子驱动风疹病毒E1基因的植物表达载体。更多还原

【Abstract】 Objective:To improve the expression of Rubella virus E1 gene in transformed tomato for further studies of E1-based edible plant vaccines.Method:The 1.1kb tomato fruit-specific promoters E8 gene and the 460bp Rubella virus E1 gene and 256bp NOS gene were amplified by polymerase chain reaclion,which were inserted into the multiple cloning site of pCAMBIA1301 to obtain the plant expression vector pCAM1301E8-E1.Then,pCAM1301E8-E1 were directly introduced into Agrobacterium tumefaciens strain EHA105,the result was identified by digestion.Result:Digestion with restriction enzymes proved that all recombinant vectors had the inserts with expected length of the target fragments,and the sequencing results were correct.Conclusion:In this study,a plant expression vector containing Rubella virus E1 gene driven by fruit specific promoter E8 was successfully constructed.更多还原