pUC-T载体的构建

Construction of pUC-T Vector

目的:构建具有较高连接效率和检出效率的克隆载体。方法:在质粒pUC18的多克隆位点内引入特异的DNA片段,酶切后得到3'末端具有突出T碱基的线性化pUC-T载体。结果:经连接效率实验,此载体具有很高的重组率。结论:成功获得了pUC-T载体;利用了质粒pUC18的多克隆位点,为亚克隆提供了丰富的酶切位点;具有蓝白斑筛选标记,提高了重组子的检出效率。

Objective To get the cloning vector with high efficiency.Methods Insert a special DNA sequence into the multiple cloning site of pUC 18 and cut the product with restriction enzyme EclHK1 to get pUC-T vector. Results Experiment of Ligation proved the high recombinational efficiency of pUC-T vector. Conclusion Get the pUC-T vector and every product of enzyme cutting has the 3′-T protruding terminus with the multiple cloning site pUC-T has lots of choices in the subcloning strategy blue/white selection enhance the efficiency of the recombinant scanning.