Highly efficient generation of T-DNA insertion lines and isolation of flanking sequence tags (FSTs) of Brachypodium distachyon
文献类型：期刊论文
| 作者 | Han, Hongjiang2; Shen, Guoan; An, Tianyue1; Song, Bo; Zhao, Suzhen; Qi, Xiaoquan
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| 刊名 | PLANT BIOTECHNOLOGY REPORTS
 |
| 出版日期 | 2018 |
| 卷号 | 12期号:4页码:237-248 |
| 关键词 | Brachypodium distachyon Agrobacterium-mediated transformation T-DNA insertion Flanking sequence tag (FST) Modified iPCR |
| ISSN号 | 1863-5466 |
| DOI | 10.1007/s11816-018-0489-4 |
| 文献子类 | Article |
| 英文摘要 | Brachypodium distachyon (Brachypodium) has been developed as a model system for the temperate grasses. Hence, establishing a large insertion mutant population and identifying T-DNA insertion sites are imperative for functional genomics studies. Currently, Brachypodium has an established T-DNA collection resource, but it is far less than needed. In addition, the conventional methods for the isolation of flanking sequence tags (FSTs) characterizing the T-DNA inserts, such as TAIL-PCR, adapter ligation-mediated PCR, and inverse PCR, are time-consuming, laborious, and expensive. In this study, Agrobacterium-mediated transformation system was optimized to generate T-DNA mutants. Approximately 7000 T-DNA insertion lines were obtained. Furthermore, a simple and highly efficient method was developed to isolate T-DNA flanking sequence tags (FSTs). The procedures simplified the multi-step reaction that is required for the conventional inverse PCR and several reactions were conducted within a microscale reaction system in one tube. It is flexible to isolate FSTs for either individual or large numbers of T-DNA lines. To rapidly process the large-scale sequence data, a serial of Perl scripts were developed using the Perl Programming Language . Using these methods, a total of 794 flanking sequences were isolated and analyzed in detail. Although the methods were developed in the process of isolating and analyzing T-DNA flanking sequences of Brachypodium, it can be applied to other plants. |
| 学科主题 | Biotechnology & Applied Microbiology ; Plant Sciences |
| 出版地 | NEW YORK |
| 电子版国际标准刊号 | 1863-5474 |
| WOS关键词 | AGROBACTERIUM-MEDIATED TRANSFORMATION ; TRANSGENE STRUCTURES ; FUNCTIONAL GENOMICS ; ARABIDOPSIS GENOME ; REVERSE GENETICS ; TAGGING LINES ; PLANTS ; RICE ; INTEGRATION ; MUTAGENESIS |
| 语种 | 英语 |
| WOS记录号 | WOS:000439478500001 |
| 出版者 | SPRINGER |
| 资助机构 | National Natural Science Foundation of ChinaNational Natural Science Foundation of China (NSFC) [31270413] |
| 源URL | [http://ir.ibcas.ac.cn/handle/2S10CLM1/20608]  |
| 专题 | 中科院植物分子生理学重点实验室 |
| 作者单位 | 1.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 2.Chinese Acad Sci, Inst Bot, Key Lab Plant Mol Physiol, Beijing 100093, Peoples R China 3.Jinan Univ, Biotechnol Inst Chinese Mat Med, Guangzhou 510632, Guangdong, Peoples R China |
| 推荐引用方式 GB/T 7714 | Han, Hongjiang,Shen, Guoan,An, Tianyue,et al. Highly efficient generation of T-DNA insertion lines and isolation of flanking sequence tags (FSTs) of Brachypodium distachyon[J]. PLANT BIOTECHNOLOGY REPORTS,2018,12(4):237-248. |
| APA | Han, Hongjiang,Shen, Guoan,An, Tianyue,Song, Bo,Zhao, Suzhen,&Qi, Xiaoquan.(2018).Highly efficient generation of T-DNA insertion lines and isolation of flanking sequence tags (FSTs) of Brachypodium distachyon.PLANT BIOTECHNOLOGY REPORTS,12(4),237-248. |
| MLA | Han, Hongjiang,et al."Highly efficient generation of T-DNA insertion lines and isolation of flanking sequence tags (FSTs) of Brachypodium distachyon".PLANT BIOTECHNOLOGY REPORTS 12.4(2018):237-248. |
入库方式： OAI收割
来源：植物研究所
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