杨树桑黄查尔酮异构酶基因克隆鉴定与表达分析  

Cloning and Expression Analysis of Chalcone Isomerase Gene from Sanghuangporus vaninii

作　　者：周园园 辛伽惠 李子豪 胡清秀[1] 邹亚杰[1] ZHOU Yuanyuan;XIN Jiahui;LI Zihao;HU Qingxiu;ZOU Yajie(Institute of Agricultural Resources and Regional Planning,Chinese Academy of Agricultural Sciences,Beijing 100081,China;Inner Mongolia University of Science and Technology Baotou Teachers′College,Baotou 014000,Inner Mongolia,China)

机构地区：[1]中国农业科学院农业资源与农业区划研究所,北京100081 [2]内蒙古科技大学包头师范学院,内蒙古包头014000

出　　处：《食用菌学报》2022年第4期10-16,共7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)查尔酮异构酶基因(SvCHI)进行克隆和生物信息学分析,利用荧光定量PCR方法研究SvCHI在菌丝体、一年生和三年生子实体中的表达情况,并测定查尔酮异构酶(SvCHI)活性。结果表明:SvCHI含有873 bp开放阅读框,编码290个氨基酸;SvCHI相对分子质量为32170,理论等电点(pI)为10.22,不稳定系数为49.47,总平均亲水性GRAVY值为-0.172;SvCHI为非跨膜蛋白,定位于线粒体,无信号肽,含有1个保守的CHI-3结构域(25~278 aa);SvCHI二级结构中无规卷曲、α螺旋、延伸链占比分别为50.69%、32.41%、16.90%;SvCHI氨基酸序列与暴马桑黄(S.baumii)和红皮孔菌(Pyrrhoderma noxium)查尔酮异构酶的相似性较高;菌丝体中SvCHI表达量显著低于一年生和三年生桑黄子实体,菌丝体中SvCHI活性显著高于一年生和三年生桑黄子实体。研究结果为进一步探讨杨树桑黄中黄酮类化合物的生物合成提供参考。The chalcone isomerase gene of Sanghuangporus vaninii(SvCHI)was cloned,analyzed for bioinformatics characteristics,and then measured for its expression and SvCHI activity in mycelium,fruiting bodies developed on artificial substrate(one year)and fruiting bodies developed on wood logs(three years),respectively.The results showed that the SvCHI gene contained an open reading frame(ORF)of 873 bp,encoding 290 amino acids.The SvCHI protein had a relative molecular weight of 32170,a theoretical isoelectric point(pI)of 10.22,an instability coefficient of 49.47,and an overall average hydrophilic GRAVY value of-0.172.SvCHI was found to be a non-transmembrane protein without signal peptide and localized in mitochondria.It contained a conserved CHI-3 structural domain at 25-278 aa.In the secondary structure of SvCHI,random coil,α-helix and extended strand accounted for 50.69%,32.41%and 16.90%,respectively.The amino acid sequence of SvCHI was found to be highly similar to that of chalcone isomerase from S.baumii and Pyrrhoderma noxium.The expression of SvCHI in mycelium was significantly lower than that in fruiting bodies developed on artificial substrate and wood logs.In contrast,SvCHI activity in mycelium was significantly higher than that in fruiting bodies developed on artificial substrate and wood logs.The results of this study provided a reference for further research on flavonoid biosynthe sis in S.vaninii.

关 键 词：杨树桑黄 查尔酮异构酶 基因克隆 表达特性 酶活 

分 类 号：S567.3[农业科学—中草药栽培]