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一种新的DNA分子克隆方法
引用本文:黄媛,陶颖,张文露,黄爱龙,胡接力.一种新的DNA分子克隆方法[J].中国科学:生命科学,2011,41(9):722-729.
作者姓名:黄媛  陶颖  张文露  黄爱龙  胡接力
作者单位:重庆医科大学, 感染性疾病分子生物学教育部重点实验室, 重庆 400016
基金项目:国家自然科学基金(批准号: 81000732)资助项目
摘    要:DNA分子克隆是基本的分子生物学实验技术,传统的分子克隆方法大多需经过酶切链接过程,但在某些情况下,没有合适的酶切位点往往会成为阻碍克隆进行的障碍.本文描述了一种新的分子克隆方法,称为不依赖酶切和链接的分子克隆(RLIC).利用RLIC,将3种不同大小的DNA片段克隆到3种不同载体,证明了这种方法的有效性和可靠性.由于该方法不受限制性酶切序列限制,省去了酶切连接步骤,因此具有很大的灵活性和简便性,在分子生物学研究方面有广泛应用前景.

关 键 词:DNA  分子克隆  方法

A New Method for DNA Molecular Cloning
HUANG Yuan,TAO Ying,ZHANG WenLu,HUANG AiLong,HU JieLi.A New Method for DNA Molecular Cloning[J].Scientia Sinica Vitae,2011,41(9):722-729.
Authors:HUANG Yuan  TAO Ying  ZHANG WenLu  HUANG AiLong  HU JieLi
Institution:Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Chongqing Medical University, Chongqing 400016, China
Abstract:DNA molecular cloning is an extensively used technique in molecular biology research. Traditional methods for molecular cloning usually include restriction digestion and ligation process. In many cases, however, cloning of a DNA fragment may be difficult because no appropriate restriction site can be found. In the present study, we have developed a new method for DNA molecular cloning, named Restriction digestion and Ligation-Independent Cloning (RLIC). We cloned three DNA fragments with different sizes into three different vectors by using RLIC, demonstrating the effectiveness of this method. Freeing from the limitation of restriction enzyme sites and omitting the ligation process, RLIC possesses great flexibility and may find broad applications in molecular biology research.
Keywords:DNA  molecular cloning  method
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