基于Xcm I酶切的pUC19-T载体的构建  

Construction of A pUC19-T Vector Based on Xcm I

作　　者：孙程龙[1,2] 李业伟[1,2] 王颖[2] 宫婷[2,3] 扈荣良[2] 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062 [2]军事医学科学院军事兽医研究所,吉林长春130122 [3]吉林农业大学动物科技学院,吉林长春130122

出　　处：《安徽农业科学》2011年第17期10182-10184,共3页Journal of Anhui Agricultural Sciences

摘　　要：[目的]构建以pUC19质粒为基础可利用XcmI内切酶制备的T载体。[方法]化学合成2条含有双XcmI酶切位点的互补寡聚核苷酸链,经过变性、复性后克隆入pUC19质粒的HindIII和BamHI位点之间,通过XcmI酶切后得到一个线性化的带有3'末端突出一个T碱基的T载体。[结果]经TA克隆验证,制备的pUC19-HB-T载体对PCR产物的克隆率达到95%以上。[结论]构建的pUC19-HB-T载体可用于PCR产物的克隆、测序及后续分子生物学操作。[ Objective] The paper aimed to construct a new T vector based on plasmid pUC19 digested with Xcm I. [ Method] Two comple- mentary oligonucleotide chains containing two Xcm I sites were synthesized. After denaturation and renaturation, the adaptor was cloned into plasmid pUCI9 between the Hind HI and BamH I sites. The new plasmid, pUC19-HB-T vector, was digested with Xcm I to derive a T-vector with 3＇ end overhanging a T base. [ Result] The constructed pUC19-I-IB-T vector was efficient in Cloning PCR products, with an efficiency of 95% at least. [ Conclusion] A new Xcm I-based pUCI9-HB-T vector was constructed, which could be applied to cloning of PCR products and other microbiology onerations.

关 键 词：T载体 Xcm I限制性内切酶 TA克隆 

分 类 号：S132[农业科学—农业基础科学]