Mi基因过表达载体和16D10基因RNAi载体的构建及转化

Construction of Mi Gene Over-Expression Vector and 16D10 Gene RNAi Vector and Their Transformation

文中采用PCR技术从番茄中克隆获得了Mi基因,采用反转录PCR(RT-PCR)技术从来源于红橘(Citus reticulate)的根结线虫中克隆获得了16D10基因,采用DNA重组技术构建了Mi基因的过表达载体和16D10基因的RNAi载体,分别命名为pB-Mi和pB-16DR.通过根瘤农杆菌介导的遗传转化,获得了经PCR检测为阳性的沙田柚转基因植株.这一结果将为研究Mi基因过表达和16D10基因的RNA干扰对柑橘根结线虫病的抗性影响提供试材和技术支持.

Mi gene was cloned from tomato with PCR amplification and 16 D10 gene was cloned from root knot nematode in Citus reticulate with reverse-transcription PCR（RT-PCR） amplification. The Mi gene over-expression vector（named pB-Mi） and 16 D10 gene RNAi vector（named pB-16 DR） were constructed with DNA molecular recombination technology. Putative transgenic plantlets,which were confirmed by PCR analysis,were obtained through genetic transformation mediated by Agrobacterium tumefaciens. The results can provide material and technical basis for exploring the effect of the over-expression of Mi gene and RNA interference of 16 D10 gene on the resistance of Citrus root knot nematode disease.