多年生簇毛麦PSY基因的克隆与进化分析
首发时间:2011-01-17
摘要:多年生簇毛麦是小麦族的珍稀物种,也是小麦染色体工程育种的重要基因源之一。根据已发表的小麦族植物PSY 基因序列的保守区设计引物,通过PCR扩增获得了多年生簇毛麦PSY基因的全序列,该基因全长为2960bp,包含6个外显子和5个内含子,被命名为DbPSY1。序列比对分析表明DbPSY1与小麦族其它植物的PSY基因有较高的同源性。将DbPSY1与禾谷类植物PSY 基因进行遗传聚类分析,结果表明,DbPSY1与偃麦草以及大麦的PSY基因亲缘关系最近而与小麦相对较远。根据序列比对结果设计多年生簇毛麦DbPSY1基因特异引物,建立了可以用于跟踪携带PSY基因的多年生簇毛麦染色体片段的分子标记,为多年生簇毛麦遗传物质向小麦的转移后鉴定提供了新的检测手段。
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Cloning and Evolutionary analysis of Dasypyrum breviaristatum PSY gene
Abstract:As a unique Triticeae species, Dasypyurum breviaristatum was important sources for wheat breeding by chromosome manipulation. In this research, PCR was carried out on the genomic DNA from Dasypyrum breviaristatum using the conserved primers specific for wheat Phytoenesynthase (PSY) genes. The PCR product was cloned and a 1960bp fragment was obtained. The sequence was predicted to have 6 exons and 5 introns. BLAST search showed that the sequence has high homology to cereal PSY gene family, and was named DbPSY1 gene. The phylogenetic analysis indicated that the DbPSY1 sequence was closed to the Thinopyrum PSY genes. In addition, on the basis of the DbPSY1 gene sequences, a pair of AS-PCR primer was designed. PCR results indicated that the AS-PCR primer allows to specifically amplifying the D. breviaristatum chromatin introduced in wheat background in wheat-Dasypyrum introgression lines.
Keywords: Dasypyrum breviaristatum PSY gene Gene cloning Specific PCR
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