| 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
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| 2014-09-22 | |
| 发表期刊 | PLOS ONE
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| ISSN | 1932-6203 |
| 卷号 | 9期号:9页码:8893-8899 |
| 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 |
| 学科领域 | Multidisciplinary Sciences |
| DOI | 10.1371/journal.pone.0107907 |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000343679800056 |
| 引用统计 | |
| 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 |
| 推荐引用方式 GB/T 7714 | 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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