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采用同源克隆、染色体步移和RT-PCR技术,首次克隆到苦荞查尔酮合酶基因(CHS)的全长DNA序列和cDNA开放阅读框(ORF)序列.序列分析表明,苦荞CHS DNA序列(GU172165)全长1 632 bp,含1个445 bp的内含子;cDNA编码区(HM852753)全长1 188 bp,编码395个氨基酸,命名为FtCHS.生物信息学分析表明,FtCHS和推导的氨基酸序列与其它植物CHS基因同源率在95%以上,含有CHS多基因家族的标签序列(GFGPG)、活性位点、底物结合口袋位点和环化反应口袋位点.半定量RT-PCR分析苦荞花期FtCHS空间表达模型表明,其表达量未成熟种子>叶>茎>花>根>成熟种子,与苦荞芦丁含量的分布基本一致,具有组织特异性。
The full length DNA sequence and cDNA open reading frame (ORF) sequence of Chalcone chalcone synthase gene (CHS) were cloned by homologous cloning, chromosome walking and RT-PCR. Sequence analysis showed that CHS The full-length DNA sequence (GU172165) was 1 632 bp in length and contained a 445 bp intron. The full-length cDNA was 1 188 bp in length encoding a 395 amino acid protein and named FtCHS. Bioinformatics analysis showed that FtCHS and The deduced amino acid sequence shared more than 95% homology with other plant CHS genes and contained the gene sequence of multiple gene family (GFGPG), active site, substrate-bound pocket site and cyclization pocket site.Semi-quantitative RT The results showed that the expression of FtCHS at the flowering stage of buckwheat showed that the expression level of immature seeds> leaves> stem> flower> root> mature seeds was basically the same as the distribution of rutin in tartary buckwheat with tissue specificity.