农杆菌介导的Thaumatin基因转化烟草的研究  

Transformation of Thaumatin Gene into Tobacco through Agrobacterium Mediation

作　　者：崔波[1] 蒋素华[1] 田云芳[1] 武振江[2] 袁秀云[1] 马杰[1] 

机构地区：[1]郑州师范学院生物工程研究所,河南郑州450044 [2]河南农业大学生命科学学院,河南郑州450002

出　　处：《江西农业学报》2013年第7期50-52,56,共4页Acta Agriculturae Jiangxi

基　　金：河南省郑州市科技攻关资助项目(083STRF40346-2)

摘　　要：以暗培养3 d的烟草叶片为受体,以Thaumatin为目的基因,应用根癌农杆菌介导法对烟草进行遗传转化,同时以烟草叶片为材料,对影响转化的几个因素进行优化研究,结果表明:侵染20 min、美罗培南浓度为25 mg/L时有利于提高转化率;经浓度为2.0 mg/L的PPT筛选获得的抗性植株经PCR、GUS组织化学染色和Southern杂交分析鉴定,初步证明外源基因Thaumatin已整合到烟草的基因组中。Tobacco leaves under dark culture for 3 days were used as receptor, and the objective gene Thaumatin was trans-formed by Agrobactorium tumefaciens mediated genetic transformation.The transformation efficiency was affected by several factors , which were optimized.The results showed that infection time of 20 min and Meropenem of 25 mg/L were beneficial to the increase in the transformation efficiency.PPT of 2.0 mg/L was used to screen resistant plant , and the resistant plant was identified by PCR , GUS histochemical staining and Southern blot analysis .These confirmed that the exogenous gene Thaumatin had been integrated into the genome of tobacco.

关 键 词：Thaumatin基因 烟草 根癌农杆菌 遗传转化 

分 类 号：Q785[生物学—分子生物学]