农杆菌介导抗虫基因GmCry1F转化玉米的研究

Study on Insect-resistant Gene of GmCry1F Transformation in Maize by Agrobacterium-mediated

培育转基因抗虫玉米是防治玉米螟虫的有效途径之一。本研究通过对来源于苏云金芽孢杆菌Cry1F野生型基因编码区密码子优化改造获得新型抗虫基因Gm Cry1F,使用农杆菌介导的玉米幼胚遗传转化法将目的基因转入玉米材料Hi-II,最后应用PCR法与蛋白免疫学法对试验结果进行鉴定检测。结果表明:成功构建了植物表达载体CUB-ubi::Gm Cry1F-35s::bar,经光照培养分化获得到转化苗25株,其中有17株的PCR和试纸条检测均呈阳性,说明Gm Cry1F基因已经整合到玉米基因组中。这一研究结果为下一步开展转基因抗虫玉米育种工作提供了较好的研究材料,也为玉米抗虫基因工程育种研究提供参考。

The cultivation of transgenic insect-resistant maize is one of the effective ways to reduce the damage caused by corn borer. A new insect-resistant gene GmCry1F was obtained by optimizing and modifying the coding regions of wild Bacillus thuringiensis CrylF gone in this study. Then it introduced target gene into maize Hi-fi via Agrobacterium-mediated, finally idenlified the experimental results by PCR and immuno strip test. The zesults showed that the plant express vector CUB-ubi：： GmCryl F-35s： ：bar has been constructed successfully. Twen- ty-five transgenlc plants were obtained through light differentiation culture in which seventeen mumgenic plants were positive by PCR and im- muno strip test. It was demonstrated that gone GmCry1F had been integrated into the maize genome. This result provided some good research materials and reference to the transgenic insect-resistant maize breeding.