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使用与Gateway技术兼容的T载体获得入门克隆
引用本文:陈其军,安瑞,周海梦,陈珈,王学臣.使用与Gateway技术兼容的T载体获得入门克隆[J].生物化学与生物物理进展,2004,31(10):951-954.
作者姓名:陈其军  安瑞  周海梦  陈珈  王学臣
作者单位:1. 中国农业大学生物学院,植物生理学与生物化学国家重点实验室,北京,100094;清华大学生物科学与技术系,北京,100084
2. 中国农业大学生物学院,植物生理学与生物化学国家重点实验室,北京,100094
3. 清华大学生物科学与技术系,北京,100084
基金项目:国家重点基础研究发展规划项目(973)(G1999011700)和国家自然科学基金资助项目(30300025).
摘    要:与Gateway技术兼容的农杆菌双元载体系统已开始应用于植物功能基因组的研究,但应用这些载体系统的一个瓶颈问题,是如何简单、经济和高效地将PCR产物或其他来源的目的DNA片段构建到入门载体上获得入门克隆.为此,将传统的TA克隆技术与Gateway重组克隆技术进行整合,构建了与Gateway技术兼容的两种TA克隆载体,用于在克隆PCR产物或其他来源的目的DNA片段的同时获得入门克隆.利用兼容Gateway技术的TA克隆载体有效地解决了上述瓶颈问题.

关 键 词:Gateway克隆技术,TA克隆技术,入门载体,T载体,入门克隆
收稿时间:2004/5/13 0:00:00
修稿时间:2004/7/30 0:00:00

Making Entry Clones Using T Vectors Compatible With The Gateway Cloning
CHEN Qi-Jun,AN Rui,ZHOU Hai-Meng,CHEN Jia and WANG Xue-Chen.Making Entry Clones Using T Vectors Compatible With The Gateway Cloning[J].Progress In Biochemistry and Biophysics,2004,31(10):951-954.
Authors:CHEN Qi-Jun  AN Rui  ZHOU Hai-Meng  CHEN Jia and WANG Xue-Chen
Affiliation:State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing 100094, China;Department of Biological Science and Biotechnology, Tsinghua University, Beijing 100084, China;State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing 100094, China;Department of Biological Science and Biotechnology, Tsinghua University, Beijing 100084, China;State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing 100094, China;State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing 100094, China
Abstract:A Gateway-compatible Agrobacterium sp. binary vector system for high-throughput functional analysis of genes in plants has been produced, but a bottleneck problem using this system for fast and reliable DNA cloning is how to obtain entry clones for the PCR products or other DNA fragments simply, economically and efficiently. To address this problem, the traditional TA cloning and the Gateway recombinant cloning techniques are integrated, and two kinds of TA cloning vectors compatible with the Gateway cloning are constructed. The vectors can be used for cloning PCR products or other DNA fragments and at the same time making entry clones in a simple, economical and efficient way.
Keywords:Gateway cloning  TA cloning  entry vectors  T vectors  entry clones
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