小反刍兽疫转基因苜蓿植物疫苗表达载体的构建

Yang Guofeng,Construction of the Transgenic Alfafa Plant Vaccine Vector of PRRV

为预防小反刍兽疫,生产稳定高效的小反刍兽疫植物疫苗,本研究以小反刍兽疫病毒(PPRV)的F基因为目的基因,以p BI121为基本表达载体,绿色荧光蛋白基因(GFP基因)为标记基因,并通过添加核基质附着区序列(MAR序列)、驻内质网膜信号序列(KDEL序列),使用强启动子Cs VMV替换启动子Ca MV35S,以期使F基因在苜蓿中能够高效表达,并最终实现预防小反刍兽疫的目的。本实验最终成功构建了五条载体:p BI121-Cs VMV-GFP-F-MAR12-MAR34、p BI121-Cs VMV-GFP-F-KDEL-MAR12-MAR34、p BI121-Cs VMVF-MAR12-MAR34、p BI121-Cs VMV-F-KDEL-MAR12-MAR34、p BI121-F。

In or der to prevent Peste des petits ruminants and produce transgenic alfalfa plant vaccine, p BI121 was chosen as the basic vector, the F and F-KDEL were used as target gene, GFP gene was employed as a marker gene. In addition, matrix attachment region(MAR) sequence and KDEL sequence were integrated and the promoter Cs VMV replaced promoter Ca MV35 S as well in order to highly express the F gene in the alfalfa. To finally achieve the purpose of preventing Peste des petits ruminants. Five expression vectors were successfully constructed. They were: p BI121-Cs VMV-GUS-GFP-F-MAR12-MAR34, BI121-Cs VMV-GUS-GFP-F-KDELMAR12-MAR34, p BI121-Cs VMV-GUS-F-MAR12-MAR34, p BI121-Cs VMV-GUS-F-KDEL-MAR12-MAR34,and p BI121-F in this study.