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当前位置: 首页 > 文章 > 三色堇花色素合成途径中部分结构基因的克隆及生物信息学分析 西北农林科技大学学报(自然科学版) 2015,43 (7) 133-142
Position: Home > Articles > Cloning and bioinformatics analysis of three structural genes involved in anthocyanin synthesis of Viola×wittrockiana Journal of Northwest A & F University(Natural Science Edition) 2015,43 (7) 133-142

三色堇花色素合成途径中部分结构基因的克隆及生物信息学分析

作  者：
龚胜;孙海燕;张珂;曾媛;李婧;王健;李新国
单  位：
海南大学园艺园林学院热带作物种质资源保护与开发利用教育部重点实验室
关键词：
三色堇;查尔酮合成酶(CHS);查尔酮异构酶(CHI);黄烷酮-3-羟化酶(F3H);生物信息学;基因克隆
摘  要：
【目的】对三色堇花色素合成的3个结构基因VwCHS、VwCHI和VwF3 H进行克隆以及生物信息学分析。【方法】利用RACE/PCR技术克隆得到VwCHS、VwCHI和VwF3 H基因的全长序列,并通过在线分析软件对其生物信息学进行分析。【结果】获得VwCHS、VwCHI和VwF3 H的cDNA全长分别是1 481,975和1 361bp;经多重序列比对发现这3个基因编码的氨基酸与其他植物相关氨基酸一致性较高,主要集中在62%~90%,并且在CHS氨基酸序列中有4个催化活性位点(Cys171、Phe222、His310、Asn343),在CHI氨基酸序列中有4个催化活性位点(Thr50、Tyr108、Asn115、Ser192),在VwF3H氨基酸序列中有5个催化活性位点(Asp218、His253、Arg215、Arg284、Ser286);VwCHS、VwCHI、VwF3H的蛋白质分子量分别为9.91,5.37,4.1ku,等电点为5.02,5.16,5.44;二级结构预测发现,VwCHS和VwCHI二级结构主要是α螺旋,分别占43.39%和41.89%,VwF3H的蛋白质二级结构主要是无规则卷曲,占42.30%;聚类分析发现VwCHS、VwCHI和VwF3H分别归属于各自的聚类簇。【结论】VwCHS、VwCHI和VwF3H是不存在信号肽的非跨膜蛋白。
译  名：
Cloning and bioinformatics analysis of three structural genes involved in anthocyanin synthesis of Viola×wittrockiana
作  者：
GONG Sheng;SUN Hai-yan;ZHANG Ke;ZENG Yuan;LI Jing;WANG Jian;LI Xin-guo;Key Laboratory of Protection and Development Utilization of Tropical Crop Germplasm Resources,Ministry of Education,College of Horticulture & Landscape Architecture,Hainan University;
关键词：
Viola×wittrockiana;;chalcone synthase(CHS);;chalcone isomerase(CHI);;flavanone-3-hydroxylase(F3H);;bioinformatics analysis;;gene clone
摘  要：
【Objective】Cloning and bioinformatics analysis were conducted for three genes,VwCHS,VwCHI and VwF3 H,which are involved in anthocyanin synthesis of pansy.【Method】The full-length cDNA of VwCHS,VwCHI and VwF3 H were cloned by RACE/PCR,and bioinformatics analysis was carried out through online software.【Result】The full-length cDNA of VwCHS,VwCHI and VwF3 H were 1 481,975 and 1 361 bp,respectively.Multiple sequence alignment revealed that the amino acids encoded by the three structural genes had high identity compared to other plants(62%-90%).There were four active sites(Cys171,Phe222,His310,and Asn343)in the amino acids sequences of CHS,four active sites(Thr50,Tyr108,Asn115,and Ser192)in CHI,and five active sites(Asp218,His253,Arg215,Arg284,and Ser286)in F3 H.Their molecular weights were 9.91,5.37 and 4.1ku,and their pI values were 5.02,5.16 and 5.44,respec-tively.The secondary structure prediction revealed that the secondary structures of VwCHS and VwCHI protein mainly containedαhelix with ratios of 43.39% and 41.89% respectively,while VwF3 H protein was mainly random coil with a ratio of 42.30%.Phylogenic analysis showed that each of the three proteins belonged to the corresponding protein family cluster.【Conclusion】VwCHS,VwCHI and VwF3 H were non-transmembrane proteins without signal peptide.

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