2008年9月
Self-EcoTILLING to identify single-nucleotide mutations in multigene family
PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY
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- 巻
- 92
- 号
- 1
- 開始ページ
- 24
- 終了ページ
- 29
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1016/j.pestbp.2008.05.001
- 出版者・発行元
- ACADEMIC PRESS INC ELSEVIER SCIENCE
TILLING (Targeting Induced Local Lesions IN Genomes) is a low-cost, high-throughput reverse genetic technique that employs a mismatch-specific endonuclease CEL-1 to discover induced point mutations in the genes of interest. The use of the TILLING technique to survey natural variation in genes is called ECOTILLING. Here, we report a modified EcoTILLING method for the discovery of mutations in multigene family, which we coin "Self-EcoTILLING" by using an allotetraploid Monochoria vaginalis ALS multigene family as an example. The mutations could be detected by TILLING of PCR products resulting from the primers specific to both Als1 and Als3 without involving the experimental step of mixture of reference and query DNA. Either of the two co-amplified loci could serve as reference DNA to the other. We demonstrate with this example that Self-EcoTILLING is a fast, reliable and economical technique of detecting single-nucleotide mutations in polyploid plants containing multigene family. (C) 2008 Elsevier Inc. All rights reserved.
- リンク情報
- ID情報
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- DOI : 10.1016/j.pestbp.2008.05.001
- ISSN : 0048-3575
- eISSN : 1095-9939
- Web of Science ID : WOS:000258560900005