摘要
启动子陷阱技术是克隆未知基因启动子的有效途径。首先用反向无启动子的GUSA替换pCAMBIA-2301载体上的GUSA及启动子序列,构建橡胶树启动子陷阱载体pCAMBIA2301:GUS-NOS,然后转化橡胶树。结果表明:获得了GUS染色阳性胚状体6个,再生后获得2个转基因株系。其中1个株系在胚状体、叶片和根中能检测到GUS基因的强烈表达,另1个株系仅在根中检测到微弱的GUS基因表达,说明获得的转基因植株为不同启动子启动GUS基因的株系。因此通过橡胶树启动子陷阱载体的构建及其在橡胶树中的表达,能够发现和克隆橡胶树内源基因的启动子。
Promoter trapping is an effective method for finding and cloning of novel promoters. Herein, the vector of promoter trapping for Hevea brasiliensis was firstly conducted by overlapping PCR, and then the transformation was carried out using the vector, finally, six transformed embryos were obtained, of which two were successfully propagated by secondary embryogenesis in two successive cycle and generated 2 transgenic lines. For one transgenic line, GUS gene was strongly expressed in the embryo, leaf and root and weak expression could be detected in the root of another transgenic line, indicating that the upstream of T-DNA exited promoter and promoter trapping could be used for finding and cloning the promoters of novel H.brasiliensis gene.
出处
《热带作物学报》
CSCD
北大核心
2013年第8期1473-1477,共5页
Chinese Journal of Tropical Crops
基金
国家天然橡胶产业技术体系(No.nycytx-34)
中央级科研院所基本科研业务费(No.1630022012006)
关键词
橡胶树
启动子
启动子陷阱
遗传转化
GUS基因表达
Hevea brasliensis
Promoter
Promoter trapping
Genetic transformation
GUS gene expression
