利用微束激光穿刺法将菜豆几丁质酶基因导入油菜的研究  

Studies on Introduction of Bean Chitinase Gene into Brassica napus via Laser Microbeam Puncture

作　　者：杨仲毅[1] 王兰岚[1] 宋桂英[1] 徐正平[1] 张丽华[1] 刘桂珍[1] 陈正华[1] 

机构地区：[1]中国科学院遗传研究所

出　　处：《激光生物学报》1999年第1期8-11,共4页Acta Laser Biology Sinica

基　　金：国家自然科学基金

摘　　要：本文以甘蓝型“双低”油菜新品种Ｈ１６５为实验材料，以子叶柄为外植体，研究了微束激光转化系统中的一些关键因素：如取材时间以播种后４—５天为宜；高渗液预处理２０—４０分钟者最佳；外源ＤＮＡ浓度以０．０５—０．１μｇ／μｌ获得转基因植株最有利等等。已建立一种操作简便，重复性好，并能准确定位于靶体细胞的转化体系。并用这个转化体系成功地将菜豆几丁酶基因导入油菜，并获得转基因植株，经ＰＣＲ扩增检测为阳性，实验结果表明外源菜豆几丁酶基因确已整合到油菜基因组中。In this research, a double low' variety of Brassica napus, H165 was used as experimental material, and cotyledon petioles were harvested as explants to study the effects of some key factors in laser microbeam transformation system. Consequently, it was found that the best time for harvest and hypertonic pretreatment are at 4-5 days after seeds' germination and 20 -40min respectively. The optimum concentration of foreign DNA was 0.1-0.05μg/μl. The transformation system which is easy to operate, repeat, and locate laser microbeam precisely on target cells has been established. Furthermore, based on this transformation system, bean chitinase gene was introduced into Brassica napus successfully. The transgenic plants which were confirmed by PCR analysis were obtained also.

关 键 词：微束激光 油菜 几丁质酶基因 

分 类 号：S562.035[农业科学—作物学] Q78[生物学—分子生物学]