白菜转脂蛋白基因CaMBP10的RNAi沉默体系的初步建立

Prelimilary Establishment of RNAi-mediated Lipid Transfer Protein Gene CaMBP10 Silencing System in Chinese cabbage

为研究白菜转脂蛋白——CaMBP10的体内功能,构建含有该基因片段的RNAi质粒,以农杆菌介导法对大白菜‘津育60’进行遗传转化,通过对转基因植物在生物与非生物胁迫中表型变化的研究,确定其体内功能。文章报告RNAi质粒的构建、鉴定及其转化植株基因组DNA中外源片段的鉴定。DNA分析结果显示,外源基因已成功插入了白菜基因组,RT-PCR检测结果显示转化植株内源CaMBP10基因的表达明显降低,为后续的转脂蛋白体内功能研究奠定了基础。

In this paper,a RNAi approach was used in down-regulated CaMBP10 gene expression to study its functions in vivo.A 273 bp fragment of CaMBP10 cDNA was cloned from Chinese cabbage＇JINYU 60＇and inserted into the RNAi vector PTCK303.According to the results of detections,sense and anti-sense of CaMBP10 fragments had been cloned into RNAi victor between the left and right of an intro from rice.The Chinese cabbage＇JINYU 60＇was transformed byAgrobacterium tumefaciens LBA4404 which harbored this anti-sense CaMBP10 gene.The result of RT-PCR analysis shows that the CaMBP10 gene expression was reduced in transgenic plants.This work would serve as a good base for following research.