Construction of a pUC19-T Vector Based on Xcm Ⅰ  

一种基于Xcm Ⅰ酶切的pUC19-T载体的构建(摘要)(英文)

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]吉林农业大学畜牧兽医学院,吉林长春130122

出　　处：《Agricultural Science & Technology》2011年第3期363-365,共3页农业科学与技术（英文版）

摘　　要：[Objective] The paper aimed to construct a new T vector based on plasmid pUC19 digested with Xcm Ⅰ. [Method] Two complementary oligonucleotide chains containing two Xcm Ⅰ sites were synthesized. After denaturation and renaturation,the adaptor was cloned into plasmid pUC19 between the Hind Ⅲ 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-HB-T vector was efficient in cloning PCR products,with an efficiency of 95% at least. [Conclusion] A new Xcm Ⅰ-based pUC19-HB-T vector was constructed,which could be applied to cloning of PCR products and other microbiology operations.[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]所构建的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。

关 键 词：T vector Xcm Ⅰ TA cloning 

分 类 号：Q78[生物学—分子生物学]