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以‘泰山红’石榴果实为试材,利用RT-PCR技术克隆石榴果皮DAHPS部分基因并对其进行了生物信息学分析。结果表明:获得1 373bp的DAHPS基因cDNA片段编码393个氨基酸,其氨基酸序列与甜橙(XP_006472576.1)、白梨(XP_009372963.1)、葡萄(NP_001268096.1)、苹果(XP_008386115.1)的相似性分别为94%、93%、92%、92%;预测蛋白质理论分子量约为44kDa,等电点为9.00,具有Cys-Ser-Ala-His保守区和典型的DAHP_synthe_2超家族保守结构域,为亲水性和跨膜蛋白,不是分泌蛋白;蛋白质二级结构主要由无规则卷曲(40.97%)、α?螺旋(38.17%)、伸展链(12.47%)和β?转角(8.40%)组成。GenBank登录号为KU198932。该研究结果为揭示石榴果实中没食子酸的代谢机理奠定了理论基础。
The ’Taishanhong’ pomegranate fruit was used as experimental material, and some genes of DAHPS in pomegranate peel were cloned by RT-PCR and analyzed by bioinformatics. The results showed that the cDNA fragment of 1 373 bp DAHPS gene encoded 393 amino acids. The deduced amino acid sequence of DAHPS gene was identical with that of orange (XP_006472576.1), white pear (XP_009372963.1), grape (NP_001268096.1), apple (XP_008386115.1) The predicted protein has a theoretical molecular weight of about 44 kDa and an isoelectric point of 9.00. The predicted protein has a conserved region of Cys-Ser-Ala-His and a typical conserved domain of DAHP_synthe_2 superfamily, with a similarity of 94%, 93%, 92% and 92% Is a hydrophilic and transmembrane protein that is not a secreted protein. The secondary structure of the protein consists mainly of random curl (40.97%), α helix (38.17%), stretch chain (12.47%) and β turn (8.40%) . GenBank accession number is KU198932. The results of this study laid the theoretical foundation for revealing the metabolic mechanism of gallic acid in pomegranate fruit.