IRDL Cloning: A One-Tube, Zero-Background, Easy-to-Use, Directional Cloning Method Improves Throughput in Recombinant DNA Preparation
Wang, Jiancai1,2; Xu, Ronghua2; Liu, Aizhong2,3; Xu,RH (reprint author),Chinese Acad Sci,Xishuangbanna Trop Bot Garden,Key Lab Trop Plant Resources & Sustainable Use,Kunming,Peoples R China.; ronghua.xu08@gmail.com; aizhongliu@xtbg.ac.cn
2014-09-22
发表期刊PLOS ONE
ISSN1932-6203
卷号9期号:9页码:8893-8899
产权排序第二
摘要Rapid and efficient construction of expression vectors and subsequent transformation are basic recombinant methods for the investigation of gene functionality. Although novel cloning methods have recently been developed, many laboratories worldwide continue to use traditional restriction digestion-ligation methods to construct expression vectors owing to financial constraints and the unavailability of appropriate vectors. We describe an improved restriction digestion-ligation (IRDL) cloning method that combines the advantage of directional cloning from double digestion-ligation with that of a low background observed by using a positive selection marker gene ccdB to facilitate digestion and ligation in a single tube. The IRDL cloning overcomes the time-consuming and laborious limits of traditional methods, thereby providing an easy-to-use, low-cost, and one-step strategy for directional cloning of target DNA fragments into an expression vector. As a proof-of-concept example, we developed two yeast vectors to demonstrate the feasibility and the flexibility of the IRDL cloning method. This method would provide an effective and easy-to-use system for gene cloning and functional genomics studies.
关键词Brazilian Amazon Oil Palm China Rubber Xishuangbanna Expansion Transformation Deforestation Resilience Dynamics Land Use Deforestation Prediction Redd Sustainability Southeast Asia
资助信息National Natural Science Foundation of China [90813004, 2009312311024]; National Natural Science Foundation of China [90813004, 2009312311024]; National Basic Research Program of China (973 Program) [2011CB915503, 2009CB522303]; National Basic Research Program of China (973 Program) [2011CB915503, 2009CB522303]
学科领域Multidisciplinary Sciences
DOI10.1371/journal.pone.0107907
收录类别SCI
语种英语
WOS研究方向Science & Technology - Other Topics
WOS类目Multidisciplinary Sciences
WOS记录号WOS:000343679800056
引用统计
被引频次:1[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.kib.ac.cn/handle/151853/18507
专题资源植物与生物技术所级重点实验室
通讯作者Xu,RH (reprint author),Chinese Acad Sci,Xishuangbanna Trop Bot Garden,Key Lab Trop Plant Resources & Sustainable Use,Kunming,Peoples R China.; ronghua.xu08@gmail.com; aizhongliu@xtbg.ac.cn
作者单位1.Univ Sci & Technol China, Sch Life Sci, Hefei 230026, Peoples R China
2.Chinese Acad Sci, Xishuangbanna Trop Bot Garden, Key Lab Trop Plant Resources & Sustainable Use, Kunming, Peoples R China
3.Chinese Acad Sci, Kunming Inst Bot, Kunming, Peoples R China
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Wang, Jiancai,Xu, Ronghua,Liu, Aizhong,et al. IRDL Cloning: A One-Tube, Zero-Background, Easy-to-Use, Directional Cloning Method Improves Throughput in Recombinant DNA Preparation[J]. PLOS ONE,2014,9(9):8893-8899.
APA Wang, Jiancai,Xu, Ronghua,Liu, Aizhong,Xu,RH ,ronghua.xu08@gmail.com,&aizhongliu@xtbg.ac.cn.(2014).IRDL Cloning: A One-Tube, Zero-Background, Easy-to-Use, Directional Cloning Method Improves Throughput in Recombinant DNA Preparation.PLOS ONE,9(9),8893-8899.
MLA Wang, Jiancai,et al."IRDL Cloning: A One-Tube, Zero-Background, Easy-to-Use, Directional Cloning Method Improves Throughput in Recombinant DNA Preparation".PLOS ONE 9.9(2014):8893-8899.
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