作　　者： (）;

机构地区： 广东药科大学药学院

出　　处： 《基因组学与应用生物学》 2019年第5期2137-2143,共7页

摘　　要： UGT催化的糖基转移反应对合成黄酮苷类至关重要。为探究影响布渣叶黄酮苷生物合成的糖基转移酶,本研究参考布渣叶转录组数据,设计特异性引物,采用RT-PCR法对布渣叶中的UDP-葡萄糖基转移酶(UDP-glucosyltransferase, UGT)基因MpUGT1的编码区进行克隆,并通过在线服务器分析该基因的生物信息学特性;利用实时荧光定量PCR方法检测了MpUGT1基因在布渣叶的芽、叶、枝、果、花不同部位的表达情况。结果表明,本研究克隆得到的布渣叶MpUGT1基因,其cDNA全长为1 397 bp,ORF 1 377 bp,编码458个氨基酸,GenBank登陆号为KY652922。生物信息学预测分析该基因编码蛋白质分子式为C2324H3643N603O671S18,相对分子质量为51 kD,理论等电点(PI)为5.99,不稳定系数为43.81,亲水性系数为-0.105。该蛋白没有跨膜结构域,含有UDP-葡萄糖醛酸/UDP-葡萄糖基转移酶家族保守结构域,不含信号肽,定位于叶绿体。RT-qPCR组织特异性表达分析结果表明MpUGT1在芽、叶、枝、果、花中均有表达,且叶的表达量最高。系统进化树分析显示Mp UGT1与同科植物黄麻、长蒴黄麻等同源UGT的相似度最高,均达到了约80%;上述结果为进一步研究该基因在布渣叶黄酮苷生物合成中的作用奠定了基础。 UGT catalyzed glycosylation reactions are essential for the synthesis of flavonoid glycosides. In order to explore an UDP-glucosyltransferase(UGT) that affects the biosynthesis of flavonoid glycosides in M. paniculata,this study designed specific primers based on the transcriptome data of M. paniculata, and cloned the coding region of UDP-glucosyltransferase gene MpUGT1 in M. paniculata by RT-PCR. The bioinformatics characteristics of MpUGT1 gene was analyzed through online server;and the expression of MpUGT1 gene in M. paniculata buds,leaves, twigs, fruits and flowers was detected by RT-qPCR. The results showed that the cloned MpUGT1 gene in this study was 1 397 bp with an open reading frame of 1 377 bp encoding 458 amino acids. The GenBank accession number was KY652 922. Bioinformatics analysis indicated that the gene encoding protein molecular formula was C2324 H3643 N603 O671 S18 and the relative molecular weight was 51.3 kD, its theoretical isoelectric point(PI)was 5.99 and the instability coefficient was 43.81, and the hydrophilicity coefficient was-0.105. MpUGT1 protein located in chloroplast without transmembrane domain and signal peptide, and contained the UDP-glucuronosyl/UDPglucosyltransferase family conserved domain. Tissue-specific expression analysis of RT-qPCR indicated that MpUGT1 was expressed in buds, leaves, twigs, fruits and flowers, and the highest expression level of MpUGT1 was in leaves. Phylogenetic tree analysis showed that MpUGT1 might have the high homology with homologous plants such as C. capsularis and C. olitoriu, which are both up to 80%. The result would be a foundation for the further study of the function of MpUGT1 gene in the biosynthesis of flavone glycosides in M. paniculata.

关 键 词： 布渣叶 糖基转移酶基因 黄酮苷 基因克隆 生物信息学分析

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