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.

作　　者：潘国庆[1,2] 张爽[3] 刘秀明[1] 李营[1] 张耀方[1] 李洪志[1] 李海燕[1] 李校堃[1] 

机构地区：[1]吉林农业大学生物反应器与药物开发教育部工程研究中心,长春130118 [2]吉林农业大学生命科学学院,长春130118 [3]中国人民解放军军事医学科学院军事兽医研究所,长春130062

出　　处：《生物工程学报》2010年第6期767-771,共5页Chinese Journal of Biotechnology

基　　金：国家高技术研究发展计划(863计划)生物反应器重大专项(No.2007AA100503);吉林省和长春市科技发展计划项目(Nos.20070922;06GG150);高等学校科技创新工程重大项目培育资金(No.70S018)资助~~

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。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.

关 键 词：油菜 转基因植株 植物生物反应器 角质细胞生长因子 

分 类 号：Q943.2[生物学—植物学]