【作者】 陈全家； 任茂智； 张锐； 曲延英； 郭三堆； 喻梅辉；

【Author】 Quanjia Chen Maozhi Ren Rui Zhang Yanying Qu Sandui Guol Meihun Yu(Agronomy department of Xin Jiang Agriculture University, Urumqi, 830052, China and The Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China )

【机构】 新疆农业大学农学院； 中国农业科学院生物技术研究所；

【摘要】 目前转基因抗虫棉所用的启动子是CAMV35S启动子,它在棉花的生殖器官表达量很低,要从根本上解决这一问题,就需要应用棉花生殖器官优势表达启动子来驱动杀虫基因在棉花的生殖器官中表达.以棉花品系Y18的基因组为模板,利用PCR技术,得到启动子片段,与pBI121载体上的gus报告基因融合,构建植物表达载体,命名为Parf-P-pBI121.利用花粉管通道法将质粒转化棉花,Kan检测获得13株阳性植株.通过PCR检测,证明外源的arf1仃启动子及gus基因已经整合到棉花的基因组中.对gus进行组织化学分析结果表明,arf1具有生殖器官表达特性,为实现杀虫基因在棉花蕾、花、铃等器官中的表达提供了理想的启动子.更多还原

【Abstract】 Now promoter used in transgenetic anti-insect cotton was CAMV 35 which had a low expression in productive organs of cotton. To resolve the problem, we applied predominant expression promoter to drive killing-insect gene to express in productive organs of cotton. Using genes of cotton line Y18 as template and PCR technique got promoter fragment, then the fragment synthesized with gus reporting gene on vector pBI121 to construct plant expression vector named Parf-p-pBI121. The wild type cotton was transformed by pollen tube pathway mediated method and 13 positive lines were screened by Kan resistance, PCR identification showed gus gene and arf1 promoter fragments had been intergrated into cotton genome. The results from chemical analysis of gus gene indicated arf1 had productive organ expression character and provided an ideal promoter for the expression in cotton bud, flower and boll on killing-insect genes.更多还原