提高农杆菌基因转化率方法的研究  

Approaches to Enhance Gene Transformation Rate by Agrobacterium tumefacines

作　　者：王艳[1] 贺宾[2] 李金耀[1] 曾幼玲[1] 秦丽[1] 蒋刚强[1] 张富春[1] 

机构地区：[1]新疆生物资源基因工程重点实验室新疆大学生命科学与技术学院,新疆乌鲁木齐830046 [2]新疆农业科学院经济作物研究所,新疆乌鲁木齐830000

出　　处：《生物技术》2005年第6期51-53,共3页Biotechnology

基　　金：国家"863"项目(No.2004AA227110-2);国家科技攻关项目(No.2001BA901A32)

摘　　要：通过甘蓝型油菜带柄子叶转化过程中转化受体是否进行预培处理、转化前菌液的对数浓度确定以及合适的再生筛选体系的摸索,研究了提高农杆菌基因转化效率的方法.试验结果表明:转化受体经过MD培养基3d的预培处理,用处于对数生长期OD600为0.6～0.8的菌液稀释至所需浸染浓度,浸染所需时间共培后,转入降低卡那霉素浓度的初筛分化培养基中,再经过后期较高浓度的卡那霉素继代筛选培养,大大提高了转化后卡那霉素抗性绿苗的得率.此研究不仅优化了油菜的转化体系和提高了油菜的转化效率,而且为农杆菌转化其它不同物种的受体材料提供了可指导的借鉴.Methods to increase transformation rate of genes in the Brassica napus L. were tried by pre-cultureing exphnts or not, confirming the logarithm concentrantion of bacterium and setting up the appropriate system of kan screening. The results showed if precultured explants with MD for 3 days were immerged for certain time by needed bacterium concentration which was diluted during phase of logarithm OD600 0.6 - 0.8, then cocultured and screend with lower concentration kanamycin, the rate of kan- resistant green shoots was increased greatly. The reasearch not only optimized the transformation system of rapes and improved the transformation rate, but also guide the directions for other transformed explants by Agrobacterium tumefacines.

关 键 词：带柄子叶 转化效率 预培养 对数期菌液浓度 筛选体系 

分 类 号：Q789[生物学—分子生物学]