[1]马国达,刘斯斯,满朝来,等.T-载体的构建[J].黑龙江农业科学,2009,(03):6-7.[doi:10.11942/j.isn1002-2767.2009.03.0006]
　MA Guo-da,LIU Si-si,MAN Zhao-lai,et al.Construction of T-Vector[J].HEILONGJIANG AGRICULTURAL SCIENCES,2009,(03):6-7.[doi:10.11942/j.isn1002-2767.2009.03.0006]
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T-载体的构建

《黑龙江农业科学》[ISSN:1002-2767/CN:23-1204/S] 卷: 期数: 2009年03 页码: 6-7 栏目: 出版日期: 2009-05-10
Title:
Construction of T-Vector
文章编号:
1002-2767(2009)03-0006-02
作者:
马国达1 ;  刘斯斯1 ;  满朝来2 ;  陈　亮1
(1 .黑龙江八一农垦大学生命科学技术学院, 黑龙江大庆163319 ;2.哈尔滨师范大学生命科学与技术学院,黑龙江哈尔滨150080)
Author(s):
MA Guo-da1 ; LIU Si-si1 ; MAN Zhao-lai2 ; CHEN Liang1
(1 .Biological Science and Technology College of Heilongjiang August First Land Reclamation University,Daqing ,Heilongjiang 163319 ;2 .Life Science and Technology College of Harbin Normal University ,Harbin,Heilongjiang 150080)
关键词:
T-载体; 构建; 克隆
Keywords:
T-vecto r; construction; cloning
分类号:
Q782
DOI:
10.11942/j.isn1002-2767.2009.03.0006
文献标志码:
A
摘要:
以pcDNA3.1/ V5-His 为骨架载体, 利用限制性内切酶EcoR V 得到平端化的线性载体, 在dTTP 和Taq 聚合酶作用下, 产生3’ 末端突出一个T 碱基的T-载体。目的基因与T-载体连接, 大肠杆菌转化, 提取质粒进行酶切鉴定, 结果表明构建的T-载体成功克隆了目的基因片段, T-载体构建成功。
Abstract:
A method for construction of a cloning vector (T-vector)for direct ligation with PCR products was described inthis passage.The T-vector derived from pcDNA3.1/ V5-His was the skeleton vector which was a linear vector derived by restrictionenzyme EcoR V and obtained a sing le T with 3’ overhanging by dTTP and Taq polymerase .After the ligation withdestination gene and T-vector , the transformation of E .coli, plasmids ex tracted were confirmed by restriction endonucleasedigestion, the results showed that the constructed T-vecto r successfully cloned the destination gene fragment, and the constructionof T-vector was successful .

参考文献/References:

[ 1] 　Zhou M Y,Clark S E ,Gomez-Sanchez C E.Universal cloning methodby TA S trategy [ J] .Biotechniques , 1995 ,19(1):34-35 .

[ 2] 　Marchurd D, Drumm M, Saulino A ,et al.Const ructi on of T-vectors ,arapid and general system for direct cloning of unmodified PCR products[J] .Nucleic Acids Res ,1991 ,19:1154.
[ 3] 　Cha J, BishaiI W,Chandrasegaran S .New vect ors f or direct cloning ofPCR product s[ J] .Gene, 1993, 136 :369-370 .
[ 4] 　Ichihara Y, Kurosaw a Y.Const ruction of new T vect ors for direct cloningof PCR products [ J] .Gene,1993 ,130 :153-154 .
[ 5] 　Ausubel F, Brent R,Kingston R.Short Protocols in Molecular Biologythi rd edition [ J] .John Wiley and Sons ,1995 ,1 :13-14.

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备注/Memo

第一作者简介:马国达(1974-), 男, 辽宁省建昌县人, 讲师, 博士, 从事肌肉发育相关基因功能研究。Tel:13936798236 ;E-mail:sihan1107@126 .com。

更新日期/Last Update: 2016-09-08