机构地区: 中山大学生命科学学院基因工程教育部重点实验室
出 处: 《生物技术》 2003年第6期31-33,共3页
摘 要: 为了提高共转化的效率 ,对《分子克隆》中感受态制备和CaCl2 转化法加以改良 ,发展了CaCl2 和MgCl2 介导的高效的感受态制备和转化方法 ,转化效率高达 10 8- 10 9转化子 /μgDNA ,完全可以满足各种转化实验 (包括共转化 )的要求。使用该法 ,我们成功地将一个受体质粒和供体质粒共转化入Cre菌株中 ,酶切结果证明供体质粒上的hIGF - 1(人胰岛素样生长因子 - 1)基因已经通过Cre酶介导的位点特异性重组连接到受体质粒的特定位点上。 An efficient procedure for the preparation of competent cells and CaCl_2 transformation was established by improving routine methods in order to increase co-transformation efficiency.The transformation efficiency of this method reaches to 10~8-10~9 transformants/μg DNA.It could meet the requirements of all kinds of transformation experiments.Two foreign vectors including a donor plasmid and a receiver plasmid were co-transformed into a Cre-strain using this method.Restriction digestion showed that hIGF-1 gene of donor plasmid had been ligated to specific site of receiver plasmid by site-specific recombination mediated by Cre enzyme.
领 域: [生物学]