根癌农杆菌感受态细胞的制备及保存方法研究

研究不同培养基、转化溶液、保存方法对根癌农杆菌感受态细胞转化率的影响，确定高转化率根癌农杆菌感受态细胞的制备及保存方法。结果表明，使用YEB培养基，单菌落接种培养过夜后以1∶100的比例扩大培养10.5 h，用20 mmol/L CaCl2＋80 mmol/L MgCl2溶液处理，所制备新鲜感受态细胞的转化率为1.59×10^5转化子/μg质粒DNA。浓度为7%的DMSO对新鲜感受态细胞保存效果较好。

Study on Preparation and Preservation Method of Competent Cell of Agrobacterium Tumefaciens

The effects of different mediums,transformation solutions and preservation methods on transformation rate of competent cell of Agrobacterium tumefaciens were studied,and the preparation and preservation methods of competent cell of Agrobacterium tumefaciens with high conversion rate were determined.The results showed that when the single colony was amplified and cultured at a dilution of 1∶ 100 for 10.5 h after it was overnight cultured in YEB medium,and it was treated with the solution of 20 mmol/L CaCl2+80 mmol/L MgCl2,the transformation rate of the prepared fresh competent cell was 1.59×105 transformants/μg plasmid DNA.And the preservation effect of 7% DMSO on the fresh competent cell was good.