烟粉虱内共生菌传毒相关groEL基因dsRNA载体的构建及其在烟草中的表达

Construction of dsRNA vector for a viral binding protein gene(groEL) from endosymbionts in Bemisia tabaci and its expression analysis in transgenic Nicotiana benthamianaa

烟粉虱内共生菌groEL基因编码一种与植物病毒传毒相关的GroEL蛋白.以B型烟粉虱为研究对象,利用1对特异引物,对B型烟粉虱体内的groEL基因进行PCR扩增,选取502 bp groEL基因片段构建dsRNA,并将其连接植物表达载体pBIN19,将重组质粒电激转化农杆菌菌株LBA4404.采用农杆菌浸染转化本氏烟草,PCR筛选表明,目的片段已整合至转基因植株的基因组DNA;RT-PCR结果表明,502 bp的groEL基因片段在转基因植株中被转录.

GroEL,a viral binding protein encoding by groEL gene from endosymbionts in Bemisia tabaci,is an important factor in transmission of plant viruses.In this study,502 bp fragment of groEL was amplified by polymerase chain reaction（PCR） and then ligated to the plant expression vector pBIN19.The recombination vector pBIN19：：groEL was transformed into Agrobacterium tumefaciens strain LBA4404 by electroporation.Transgenic Nicotiana benthamiana plants were acquired by Agrobacterium-mediated approach.PCR and RT-PCR were performed to analyze groEL＇s transformation and expression in transgenic plants,respectively.The results indicated that groEL gene was not only integrated into chromosome DNA of transgenic N.benthamiana,but also transcribed in transgenic plants successfully.