β-伴大豆球蛋白α'亚基和β亚基基因双价RNAi表达载体抑制大豆抗原蛋白的表达

Expression of inhibited antigenic soybean protein by RNAi vector of soybean β-conglycinin α'and β-subunits bivalent gene

为幼龄动物提供优质的大豆饲料,同时有效抑制α＇亚基和β亚基基因表达量,根据dsRNAi原理,以α＇亚基基因RNAi重组植物表达载体pCAMBIA3301-PFNZ-BADH为基础,构建以BADH安全标记为筛选标记的α＇亚基和β亚基基因双价RNAi植物表达载体。用根癌农杆菌介导法转化大豆子叶节,并对再生植株进行分子生物学检测。结果表明：成功构建β-伴大豆球蛋白α＇亚基和β亚基基因双价RNAi表达载体,并获得大豆转基因植株。T1转基因大豆植株经PCR检测得到11株阳性转化株,经Southern杂交检测证明构建的双价植物表达载体已经以1-2个拷贝整合到大豆基因组中,实时荧光定量PCR检测表明β-伴大豆球蛋白α＇亚基和β亚基基因的表达被明显抑制。

In order to provide better soybean forage for young animal,RNAi expression vector of soybean β-conglycinin α＇ and β- subunits bivalent gene was built to decrease antigen quantity of soybean protein. Based on RNAi expression vector p CAMBIA3301- PFNZ- BADH,fracture of sense and anti- sense of β subunits gene was linked to each side of fracture of sense and anti- sense of α＇ subunits on p CAMBIA3301- PFNZ- BADH by digestion. Then the vector was transformed into soybeans cotyledonary node via Agrobacterium tumefaciens- mediated method. Putative transgenic soybean were analyzed by PCR. 11 transgenic soybean plants were obtained. Southern blot of T1 plants showed that the β- conglycinin α＇and β- subunits bivalent gene was successfully integrated into soybean genome. qRT- PCR in soybean seed showed that β- conglycinin α＇ subunit and β subunit were both decreased significantly.