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根据GenBank发表的丁香酚合成酶(EGS)基因的蛋白保守序列设计引物,利用RT-PCR结合RACE-PCR技术,从古老月季(Rosa chinensis)品种‘月月粉’盛开期的花瓣中获得了1个新的丁香酚合成酶基因RcEGS1,GenBank登录号为JQ522949。该基因全长为1171bp,开放阅读框(ORF)951bp,编码317个氨基酸;蛋白质理论分子量为35.64kD,等电点为7.36。氨基酸同源性分析表明,RcEGS1与矮牵牛PhIGS1的氨基酸同源性达到70%,与罗勒ObEGS1的同源性为59%,与月季RhEGS1的同源性仅为48.2%。运用实时荧光定量PCR法分析RcEGS1在不同组织部位和不同开放时期花瓣中的表达,发现其在叶片和茎段中没有表达,在花瓣和萼片中有表达,在盛开期的花瓣中表达量最高。
Primers were designed based on the conserved sequence of the eugenol synthase (EGS) gene published in GenBank. The primers were designed from the petals of Rosa chinensis blooms by RT-PCR and RACE-PCR A new eugenol synthase gene RcEGS1, GenBank accession number JQ522949. The full length of this gene was 1171bp and contained 951bp of open reading frame (ORF) encoding 317 amino acids. The theoretical molecular weight of the protein was 35.64kD and the isoelectric point was 7.36. Amino acid homology analysis showed that the amino acid homology between RcEGS1 and petunia PhIGS1 was 70%, its homology with basil ObEGS1 was 59%, and its homology with rose RhEGS1 was only 48.2%. The expression of RcEGS1 in petals at different tissue sections and in different open periods was analyzed by real-time fluorescence quantitative PCR. The expression of RcEGS1 was not observed in leaves and stem segments, but was expressed in petals and sepals, and was highest in petals at full bloom.