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Analysis and validation of genome-specific DNA variations in 5′flanking conserved sequences of wheat low-molecular-weight glutenin subunit genes

Analysis and validation of genome-specific DNA variations in 5′ flanking conserved sequences of wheat low-molecular-weight glutenin subunit genes
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摘要 The thirty-three 5′ flanking conserved sequences of the known low-molecular-weight subunit (LMW-GS) genes have been divided into eight clusters, which was in agreement with the classification based on the deduced N-terminal protein sequences. The DNA polymorphism between the eight clusters was obtained by sequence alignment,; a total of 34 polymorphic positions were observed in the approximately 200 bp regions, among which 18 polymorphic positions were candidate SNPs. Seven cluster-specific primer sets were designed for seven out of eight clusters containing cluster-specific bases, with which the genomic DNA of the ditelosomic lines of group 1 chromosomes of a wheat variety ‘Chinese Spring’ was employed to carry out chromosome assignment. The subsequent cloning; DNA sequencing of PCR fragments validated the sequences specificity of the 5′ flanking conserved sequences between LMW-GS gene groups in different genomes. These results suggested that the coding; 5′ flanking regions of LMW-GS genes are likely to have evolved in a concerted fashion. The seven primer sets developed in this study could be used to isolate the complete ORFs of seven groups of LMW-GS genes, respectively,; therefore possess great value for further research in the contributions of a single LMW-GS gene to wheat quality in the complex genetic background; the efficient selections of quality-related components in breeding programs. The thirty-three 5′ flanking conserved sequences of the known low-molecular-weight subunit (LMW-GS) genes have been divided into eight clusters, which was in agreement with the classification based on the deduced N-terminal protein sequences. The DNA polymorphism between the eight clusters was obtained by sequence alignment, and a total of 34 polymorphic positions were observed in the approximately 200 bp regions, among which 18 polymorphic positions were candidate SNPs. Seven cluster-specific primer sets were designed for seven out of eight clusters containing cluster-specific bases, with which the genomic DNA of the ditelosomic lines of group 1 chromosomes of a wheat variety ‘Chinese Spring’ was employed to carry out chromosome assignment. The subse-quent cloning and DNA sequencing of PCR fragments validated the sequences specificity of the 5′ flanking conserved sequences between LMW-GS gene groups in different genomes. These results suggested that the coding and 5′ flanking regions of LMW-GS genes are likely to have evolved in a concerted fashion. The seven primer sets developed in this study could be used to isolate the com-plete ORFs of seven groups of LMW-GS genes, respectively, and therefore possess great value for further research in the contributions of a single LMW-GS gene to wheat quality in the complex genetic background and the efficient selections of quality-related components in breeding programs.
出处 《Science China(Life Sciences)》 SCIE CAS 2006年第4期322-331,共10页 中国科学(生命科学英文版)
基金 supported by the National High Technology Research and Development Program of China(863 Program)(Grant No.2003AA207100) National Natural Science Foundation of China(Grant Nos.30300219&30571163) the Foundation for the Author of National Excellent Doctoral Dissertation of China from Ministry of Education,China(Grant Nos.200357&200458) supported by the Program for Changjiang Scholars and Innovative Research Teams in University of China(Grant No.IRT0453)
关键词 wheat LMW-GS gene 5' flanking sequence DNA polymorphism SNP GENE cloning. wheat LMW-GS gene 5′ flanking sequence DNA polymorphism SNP gene cloning
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