轮状病毒外壳蛋白VP7在转基因番茄果实中的特异表达

Specific Expression of Rotavirus Outer Capsid Protein VP7 in Transgenic Tomato Fruits

将轮状病毒外壳蛋白VP7基因克隆到含有番茄果实特异性启动子TFP的植物表达载体pTF ,并转化到根癌农杆菌 (Agrobacteriumtumefaciens)菌株EHA10 5中 ,采用叶盘转化法转化番茄 (LycopersiconesculentumMill.)栽培品种TX0 0 14 ,获得了转基因植株。经PCR、PCR Southernblot和Southernblot分析表明VP7基因已整合到转基因番茄植株的核基因组中 ,RT PCR。

Rotaviruses are the most common cause of severe diarrhea in children all over the world.Their major outer capsid protein VP7 is a primary candidate for the vaccines.Introducing VP7 into tomato to develop edible plant vaccine of rotaviruses would change the traditional means of production of vaccines and the cost of vaccine production would be reduced greatly.In this study,plant expression vector pTF/VP7 was constructed and it is under control of tomato-fruit-specific promoter (TFP).After co-cultivation with Agrobacterium tumefaciens EHA105 and several times of kanamycin-resistant selection,the regenerated tomato plantlets were analyzed further by PCR,PCR-Southern blot and Southern blot.The results confirmed that the VP7 gene had been integrated into tomato genome.RT-PCR and Western blot analysis showed further that:VP7 was expressed specifically in transgenic tomato fruits.