摘要
载体构建是建立真菌遗传转化体系的基础。本研究以pCAMBIA1305.1双元载体为基本骨架,构建棉花黄萎病菌遗传转化T-DNA质粒双元载体,通过限制性内切酶酶切、补平、连接以及去磷酸化等措施将pCAMBIA1305.1上Hygromycin抗性基因的CaMV 35S启动子替换成构巢曲霉的trpC启动子。经大肠杆菌转化后对转化子进行酶切验证,并将新构建的载体1305-3分别转入农杆菌菌株LBA4404和EHA105。为农杆菌介导的黄萎病菌遗传转化体系的建立和优化提供了存在于不同农杆菌菌株中的供试载体。
Construction of binary vector is the basis for agrobacterium-mediated transformation of fungi. Binary vector used for agrobacterium-mediated transformation of VerticiUium dahliae was constructed with pCAMBIAI305.1 as the backbone.CaMV 35S promotor of Hygromycinin pCAMBIAI305.1 was replaced by trpC promotor of aspergillus nidulans by procedures such as digestion with restriction enzyme, dephosphatasing with BCIP, and lingation with T4 lingase etc. The new constructed vector, 1305-3, was transformed to the two agrobacterium strains, LBA4404 and EHAI05, respectively. They were used to provide different types of vector in different types of agrobactefium strain for further genetic transformation of Verticillium dahliae and optimizing the transformation condition.
出处
《石河子大学学报（自然科学版）》
CAS
2007年第1期34-38,共5页
Journal of Shihezi University(Natural Science)
关键词
载体构建
农杆菌介导的转化
黄萎病菌
棉花
vector construction
agrobacterium-mediated transformation
VerticiUium dahliae
cotton
