杨树桑黄6-磷酸海藻糖合成酶基因克隆鉴定与表达分析  

Cloning and Expression Analysis of Trehalose-6-Phosphate Synthase from Sanghuangporus vaninii

作　　者：徐丛涛 李子豪 潘晋龙 李海康 胡清秀[2] 邹亚杰[2] 陈晓华 弥春霞[1] XU Congtao;LI Zihao;PAN Jinlong;LI Haikang;HU Qingxiu;ZOU Yajie;CHEN Xiaohua;MI Chunxia(College of Life Science and Technology,Mudanjiang Normal University,Mudanjiang 157000,Heilongjiang,China;Institute of Agricultural Resources and Regional Planning,Chinese Academy of Agricultural Sciences,Beijing 100081,China;Zhejiang Qianji Fang Pharmaceutical Technology Co.,Ltd.,Hangzhou 310000,Zhejiang,China)

机构地区：[1]牡丹江师范学院生命科学与技术学院,黑龙江牡丹江157000 [2]中国农业科学院农业资源与农业区划研究所,北京100081 [3]浙江千济方医药科技有限公司,浙江杭州310000

出　　处：《食用菌学报》2024年第1期45-52,共8页Acta Edulis Fungi

基　　金：国家现代农业产业技术体系(CARS-20)。

摘　　要：对杨树桑黄(Sanghuangporus vaninii)6-磷酸海藻糖合成酶基因(SvTPS)进行克隆和生物信息学分析,并进行原核表达;采用荧光定量PCR方法研究SvTPS在杨树桑黄菌丝体和不同年份子实体中的表达情况,测定菌丝体和不同年份子实体中SvTPS活性。结果表明:SvTPS DNA序列长度为1894 bp,编码576个氨基酸。SvTPS相对分子质量为65130,等电点为6.21,脂肪系数为88.14,平均亲水系数为-0.281,不稳定系数为42.41。SvTPS氨基酸序列与暴马桑黄(S.baumii)的相似性最高,仅有一个TPS单结构域;SvTPS二级结构中α-螺旋、β-折叠、延伸链、无规卷曲占比分别为48.61%、5.56%、15.62%、30.21%。三年生子实体SvTPS的表达量最高,菌丝体的表达量最低。菌丝体的SvTPS活性最低;随着栽培时间增加,SvTPS活性增加,三年生子实体的最高。研究结果为进一步探讨SvTPS在杨树桑黄生长发育过程中的功能提供参考。The trehalose 6-phosphate synthase(SvTPS)gene of Sanghuangporus vaninii was cloned,analyzed by bioinformatics and heterologously expressed in E.coli.The expression of SvTPS in mycelia and different ages of fruiting bodies was measured by fluorescence quantitative PCR,and the activity of SvTPS was also determined.The results showed that SvTPS has an open reading frame of 1894 bp,encoding 576 amino acids.The molecular weight,theoretical isoelectric point(pI),aliphatic index,grand average of hydrophilicity(GRAVY),instability index of SvTPS were estimated to be 65130,6.21,88.14,-0.281 and 42.41,respectively.The amino acid sequence of SvTPS has the highest similarity with S.baumii,having only one TPS domain.In terms of the secondary structure of SvTPS,α-helix,β-sheet,extended strand and random coil account for 48.61%,5.56%,15.62%and 30.21%,respectively.The expression level of SvTPS was the highest in three-year-old fruiting bodies and the lowest in mycelia.SvTPS activity was the lowest in mycelia,and increased with the increase of cultivation time,reaching the highest in three-year-old fruiting bodies.The results provided a reference for further exploring the role of SvTPS in the growth and development of S.vaninii.

关 键 词：杨树桑黄 6-磷酸海藻糖合成酶 基因克隆 蛋白表达 

分 类 号：S567.3[农业科学—中草药栽培]