【摘要】 在同一反应体系内,利用IIS型限制性核酸内切酶和DNA连接酶进行"Golden Gate"克隆方法,将多个目的基因片段按照设定的顺序连接,实现多基因片段一步"无缝"连接。为编辑小鼠β酪蛋白基因位点,利用"Golden Gate"克隆法,构建了小鼠β酪蛋白基因位点同源重组靶向载体。更多还原

【Abstract】 The ’Golden Gate’ cloning is based on the ability of type IIS restriction enzymes and DNA ligase enzymes to assemble multiple DNA fragments in a defined linear order in one tube and one step.The method is efficient and yields recombinant vectors that do not contain unwanted sequences in the final construct after just a short time restriction-ligation.To edit mouse β casein gene,recombination targeting vectors of the mouse β casein gene targeting locus were constructed by the ’Golden Gate’ cloning.更多还原