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题名: IRDL Cloning: A One-Tube, Zero-Background, Easy-to-Use, Directional Cloning Method Improves Throughput in Recombinant DNA Preparation
作者: Wang, Jiancai1, 2; Xu, Ronghua2; Liu, Aizhong2, 3
刊名: PLOS ONE
出版日期: 2014-09-22
卷号: 9, 期号:9, 页码:8893-8899
关键词: BRAZILIAN AMAZON ; OIL PALM ; CHINA ; RUBBER ; XISHUANGBANNA ; EXPANSION ; TRANSFORMATION ; DEFORESTATION ; RESILIENCE ; DYNAMICS ; Land use ; Deforestation ; Prediction ; REDD ; Sustainability ; Southeast Asia
学科分类: Multidisciplinary Sciences
DOI: 10.1371/journal.pone.0107907
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
文章类型: Article
英文摘要: 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.
类目[WOS]: Multidisciplinary Sciences
研究领域[WOS]: Science & Technology - Other Topics
关键词[WOS]: LIGATION-INDEPENDENT CLONING ; PLANT FUNCTIONAL GENOMICS ; TOXIN-ANTITOXIN SYSTEMS ; ONE-STEP ; PCR PRODUCTS ; POSITIVE-SELECTION ; ESCHERICHIA-COLI ; VECTOR ; TRANSFORMATION ; GENE
收录类别: SCI
项目资助者: National Natural Science Foundation of China [90813004, 2009312311024] ; National Basic Research Program of China (973 Program) [2011CB915503, 2009CB522303]
语种: 英语
WOS记录号: WOS:000343679800056
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.kib.ac.cn/handle/151853/18507
Appears in Collections:资源植物与生物技术所级重点实验室_期刊论文

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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

Recommended Citation:
Wang,Jiancai;Xu,Ronghua;Liu,Aizhong.IRDL Cloning: A One-Tube, Zero-Background, Easy-to-Use, Directional Cloning Method Improves Throughput in Recombinant DNA Preparation,PLOS ONE,2014,70(46):8893-8899
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