Simultaneous mutation detection of three homoeologous genes in wheat by High Resolution Melting analysis and Mutation Surveyor？ 英文参考文献.docVIP

BMC Plant Biology BioMedCentral Methodology article Open Access Simultaneous mutation detection of three homoeologous genes in wheat by High Resolution Melting analysis and Mutation Surveyor? Chongmei Dong*1, Kate Vincent1,2 and Peter Sharp1 Address: 1Plant Breeding Institute, University of Sydney, PMB 4011, Narellan NSW 2567, Australia and 2Australian Centre for Plant Functional Genomics, PMB 1, Glen Osmond SA 5064, Australia Email: Chongmei Dong* - chongmei.dong@.au; Kate Vincent - kate.vincent@.au; Peter Sharp - peter.sharp@.au * Corresponding author Published: 4 December 2009 Received: 21 May 2009 Accepted: 4 December 2009 BMC Plant Biology 2009, 9:143 doi:10.1186/1471-2229-9-143 This article is available from: /1471-2229/9/143 ? 2009 Dong et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract Background: TILLING (Targeting Induced Local Lesions IN Genomes) is a powerful tool for reverse genetics, combining traditional chemical mutagenesis with high-throughput PCR-based mutation detection to discover induced mutations that alter protein function. The most popular mutation detection method for TILLING is a mismatch cleavage assay using the endonuclease CelI. For this method, locus-specific PCR is essential. Most wheat genes are present as three similar sequences with high homology in exons and low homology in introns. Locus-specific primers can usually be designed in introns. However, it is sometimes difficult to design locus-specific PCR primers in a conserved region with high homology among the three homoeologous genes, or in a gene lacking introns, or if information on introns is not available. Here we describe a mutation detection method which combines