轮状病毒G3血清型抗原蛋白VP7在马铃薯中的表达

EXPRESSION OF ANTIGEN GENE VP7 FROM G3 STRAIN OF ROTAVIRUS IN POTATO

采用RT-PCR方法扩增得到轮状病毒外壳蛋白G3VP7基因,其5′端和3′端分别与马铃薯Y病毒(PVY)的前导序列和PVY 3′端的非编码区序列相连接,将拼接好的目的片段克隆到植物表达载体pB I121质粒上,利用三亲融合法将重组质粒转入农杆菌,农杆菌介导转化植株,经PCR、PCR-Southern b lot、Southern b lot和ELISA鉴定,确定VP7基因已转入马铃薯基因组,并得以正确表达,为以植物为生物反应器生产口服疫苗奠定了基础。

Antigen gene VP7 of Human rotavirus was amplified by RT-PCR, and its 5＇end and 3＇end were respectively combined with the leading sequence and 3＇-UTR of PYY. The modified gene was cloned into plant expression vector pBI121 containing CaMV35s. The plasmids recombined were transferred into Agrobacterium tumefaciens. Resistant plants of potato were produced using an Agrobaeterium- mediated transformation system. Result showed that antigen genes were transmitted into potatoes and antigen VP7 has been expressed by PCR, PCR-Southern blot, Southern blot and ELISA assay. The expression system will be useful for producing oral vaccine in the future.