暴马桑黄倍半萜合酶基因SbTps1的克隆及表达分析  

Cloning and Function Identification of the Sesquiterpenes Synthase Gene SbTps1 in Sanghuangporus baumii

作　　者：乌木提·巴合提别克 唐玉倩 王淑婷 李亚伟 邹莉[1] WUMUTI Bahetibieke;TANG Yuqian;WANG Shuting;LI Yawei;ZOU Li(School of Forestry,Northeast Forestry University,Harbin 150040,China)

机构地区：[1]东北林业大学林学院,哈尔滨150040

出　　处：《森林工程》2021年第4期33-39,46,共8页Forest Engineering

基　　金：中央财政林业科技推广项目(黑[2021]TG14号)。

摘　　要：倍半萜(Sesquiterpenes)在动植物及微生物体内广泛存在,且具有重要的生理调节功能。本研究从暴马桑黄转录组数据库中筛选出1个编码暴马桑黄萜类合酶的基因,命名为SbTps1,并对其进行克隆及表达分析,经测序该基因全长1281 bp,编码426个氨基酸,编码蛋白的相对分子质量为48.17 ku。SDS-PAGE分析结果表明,在相对分子质量69.17 ku(包含21 ku标签蛋白)附近出现与预期大小一致的蛋白条带。系统发育进化树分析表明,暴马桑黄SbTps1蛋白和嗜蓝孢孔菌Tps蛋白同源性最高;荧光定量分析结果表明,SbTps1基因转录水平在不同发育阶段有差异性,并且在菌丝发酵培养第14天时SbTps1基因转录水平达到最高状态,该结果与鲍姆纤孔菌(Inonotus baumii)倍半萜合酶基因的转录水平变化趋势相似,因此可以推测SbTps1与倍半萜合成有关,初步鉴定该基因为倍半萜合酶基因,为后续暴马桑黄倍半萜合酶基因功能的研究奠定理论基础。Sesquiterpenes are widely found in plants,animals and microorganisms,and have important physiological regulatory functions.In this study,a gene encoding terpenoid synthase was screened out from the transcriptome database of Sanghuangporus baumii,named SbTps1,and its cloning and expression were analyzed.Sequence analysis showed that the full length of the gene was 1281 bp,encoding 426 amino acids with a molecular weight of 48.17 ku.The results of SDS-PAGE showed that a protein band of the same size as expected appeared near 69.17 ku(including 21 ku labeled protein).Phylogenetic tree results showed that SbTps1 protein and TPS protein of Formitiporia mediterranea had the highest homology.Quantitative real-time PCR analysis results showed that SbTps1 gene transcription level had differences in different development stages,and the transcription level of SbTps1 gene reached the highest state on the 14 th day of mycelium fermentation culture,which was similar to the transcription level of Inonotus baumii sesquiterpene synthase gene.Therefore,it can be inferred that SbTps1 was related to sesquiterpene synthesis,and the gene was preliminarily identified as a sesquiterpene synthase gene,which laid a foundation for the further study on the function of sesquiterpene synthase gene in Sanghuangporus baumii.

关 键 词：暴马桑黄 倍半萜 倍半萜合酶基因 基因克隆与序列分析 原核表达 荧光定量 

分 类 号：S567.3[农业科学—中草药栽培]