论文部分内容阅读
本研究使用In-Fusion TM试剂盒构建植物表达载体p TF101.1-P35S::Cry1Ab-Ma,以玉米杂交种HiⅡ幼胚为受体材料,采用农杆菌介导法将P35S驱动的抗虫基因Cry1Ab-Ma基因转入玉米,经双丙氨膦筛选后的抗性愈伤分化出43株玉米幼苗,经目的基因PCR及表达蛋白检测,其中的28株呈阳性。T1代植株叶片蛋白粗提物经Cry1Ab免疫试纸条检测,结果表明目的基因表现出分离,在部分转基因植株后代中获得了表达。本研究为培育优良的抗虫玉米自交系奠定了基础。
In this study, the plant expression vector p TF101.1-P35S :: Cry1Ab-Ma was constructed by using In-Fusion ™ kit. Using the HiⅡ immature embryo as the recipient material, Agrobacterium-mediated transformation of P35S-driven insect- Cry1Ab-Ma gene was transferred into maize, and 43 maize seedlings were isolated from the resistant plants after screening by bialaphos. The target gene was detected by PCR and expressed protein, of which 28 were positive. T1 generation plant leaf protein crude extract by CrylAb immunoassay strip test results showed that the target gene showed separation, obtained in some progeny transgenic plant expression. This study laid the foundation for cultivating excellent insect-resistant maize inbred lines.