基于Cre/loxP重组系统的多基因载体构建及烟草转化研究

Construction of multi-gene expression vector and tobacco transformation based on Cre-LoxP recombination system

利用一套基于Cre/loxP重组系统的植物多基因表达载体系统将苏云杆菌毒蛋白基因(BtCryIAc)、甜菜碱醛脱氢酶基因(BADH)、GA20氧化酶基因(pttGA20ox)和rolB基因构建于植物表达载体pYL1305上,命名为pYL1305BBGR。采用农杆菌介导的叶盘法转化烟草,经PCR和Southern blot检测发现,多基因已成功转入烟草基因组中;经半定量RT-PCR检测证实,4个基因均能正常表达。

Using a system of multi-gene plant expression vector based on a set of Cre-LoxP recombination system,the BtCryIAc,BADH,pttGA20ox and rolB genes were inserted into the plant expression vector pYL1305,which was named pYL1305BBGR.The tabacco was transformed by leaf disc method mediated by Agrobacterium tumefaciens.Tested by PCR and Southern blot,the multi genes have been successfully transferred into the genome of tobacco.The semi-quantitative RT-PCR analysis showed that all the 4 genes were expressed normally.