蓝莓3-O-葡萄糖基转移酶基因克隆及生物信息学分析  

Cloning and Bioinformatics Analysis of UDP-glucose:Anthocyanidin 3-O-glucosetransferase Gene in Blueberry

作　　者：刘娜 田雨菁 冯哲瀚 李虎良 张蕾 黄金海 华德平 LIU Na;TIAN Yujing;FENG Zhehan;LI Huliang;ZHANG Lei;HUANG Jinhai;HUA Deping(School of Life Sciences,Tianjin University,Tianjin 300072,China)

机构地区：[1]天津大学生命科学学院,天津300072

出　　处：《华北农学报》2023年第2期120-128,共9页Acta Agriculturae Boreali-Sinica

基　　金：蔬菜种质创新国家重点实验室开放课题(201901);国家自然科学基金项目(31400236);天津市重大专项合作项目(17ZXZYNC00050)。

摘　　要：为研究蓝莓3-O-葡萄糖基转移酶基因(3GT)的性质和功能,探明3-O-葡萄糖基转移酶在花青素合成过程中发挥的作用,以成熟的蓝莓果实为材料,利用同源克隆获得了3GT基因的单一拷贝,运用生物信息学软件分析预测蓝莓3GT的序列特征。结果表明:3GT基因长度为1371 bp,编码456个氨基酸,位于蓝莓的4号染色体末端,含有3个外显子和2个内含子。蛋白的分子式为C_(2279)H_(3521)N_(595)O_(650)S_(15),理论等电点(pI)为6.14。信息分析预测3GT蛋白有37个磷酸化位点,亲水指数小于0为亲水性蛋白,不稳定系数大于40为不稳定蛋白,无跨膜区和信号肽;蛋白质二级结构由α-螺旋(Hh)、延伸链(Ee)、β-转角(Tt)和无规则卷曲(Cc)4种结构形式组成,其中α-螺旋占比最高,达40%;3GT属植物糖基转移酶家族(GTB-型),PSPG结构域中第44位是组氨酸,糖基供体可能为UDP-半乳糖;与猕猴桃亲缘关系最近,同源性为67%;分子对接显示蓝莓3GT与矢车菊素分子存在4种相互作用力;蓝莓3GT启动子上游可能具有与光响应、抗胁迫响应及特定激素响应相关的顺式作用元件。本研究克隆得到了蓝莓3GT基因,通过在线网站和预测软件对其进行了生物信息学分析和功能预测。In order to study the function of 3-O-glucosyltransferase gene(3GT)in blueberry and to explore the roles of 3-O-glucosyltransferase in anthocyanin synthesis,a single copy of 3GT was cloned from the cDNA of blueberry fruit by RT-PCR,which was 1371 bp and encoded 456 amino acids.3GT was located at the end of chromosome Ⅳ in blueberry and contains 3 exons and 2 introns.The molecular formula of the protein was C_(2279)H_(3521)N_(595)O_(650)S_(15),and the theoretical isoelectric point(pI)was 6.14.Information analysis predicted that 3GT was one hydrophilic protein and unstable protein with 37 phosphorylation sites,without signal peptide,or transmembrane domain.The secondary structure of protein consists of α-helix(Hh),extended chain(Ee),β-angle(Tt)and random curl(Cc),among which α-helix accounted for 40%.As one number of GTB-type glycosyltransferase family,the 44th position in the PSPG domain of 3GT was histidine,and the glycosylase donor might be UDP-galactosewas.3GT was closely related to Kiwifruit,with the homology of amino acids reached 67%.Auto-Dock showed that 3GT had four interaction forces with zymolyte of cyanidin.There were multiple cis-regulatory elements that might relate to light response,stress resistance response and specific hormone response in the promoter of 3GT.The blueberry 3GT gene was cloned.The bioinformatics analysis and function prediction were carried out through online website and prediction software to study the characteristics and function of 3GT.

关 键 词：蓝莓 3GT 基因特征分析 花青素合成 

分 类 号：Q78[生物学—分子生物学] S663.9[农业科学—果树学]