角质细胞生长因子2基因 (KGF2) 的克隆及其对油菜的遗传转化

Cloning of Keratinocyte growth factor 2 gene (KGF2) and its transformation to Brassica napus L.

摘要:

近年来，植物表达人源蛋白的研究逐渐增多。本研究以人角质细胞生长因子 (KGF2) 基因cDNA为基础，根据植物偏好密码子进行改造，并利用PCR方法合成KGF2基因全长cDNA。在此基础上构建了KGF2的植物油体系统表达载体p1390-YO-KGF2，并采用农杆菌介导法对甘蓝型油菜无菌苗的子叶进行转化。通过PCR、Southern杂交和Western blotting检测，证明外源基因KGF2已经转入油菜中并得到成功表达。

Abstract:

Recently, more research about the plant bioreactor expressing genes encoding human proteins was reported. In the present study, the cDNA of the human gene keratinocyte growth factor 2 (KGF2) was replaced with plant preferred codons by PCR, and the modified full-length cDNA was cloned into the plant expression vector pCAMBIA-YO containing the oil-body promoter. The fusion construct pCAMBIA-YO-KGF2 was transformed into Brassica napus by Agrobacterium tumefacien-mediated cotyledon transformation method. The transgenic seedlings were identified by PCR, Southern and western blot analysis all showed that KGF2 gene was successfully expressed in in transgenicBrassica napus.