摘要
为研究暴马桑黄(Sanghuangporus baumii)三萜合成途径的调控机制,对该途径中的关键酶——甲羟戊酸激酶(Mevalonate kinase,MVK)基因进行克隆及差异表达分析。以暴马桑黄cDNA为模板,采用同源克隆得到MVK基因cDNA序列全长,命名为SbMVK(登录号为MN793978)。生物信息学分析显示,SbMVK基因cDNA序列长1041 bp,编码346个氨基酸,蛋白分子量为36.6 ku,属于GHMP kinase超家族成员;系统进化树结果表明,暴马桑黄MVK蛋白和紫芝MVK蛋白同源性最高。采用qRT-PCR技术对暴马桑黄不同发育阶段SbMVK基因进行转录水平分析,并对相应阶段的三萜进行提取和含量测定,结果表明:暴马桑黄SbMVK基因转录水平和三萜含量均先升后降,并且二者变化趋势基本一致,推测SbMVK基因在暴马桑黄甲羟戊酸途径中参与三萜调控工作,此研究结果可为深入了解该基因在暴马桑黄三萜合成中的功能及获得高三萜产量的菌株提供理论依据。
To reveal the regulation mechanism of triterpenoids biosynthesis pathway in S.baumii,a mevalonate kinase(MVK)gene involved in the triterpenoids biosynthesis pathway was cloned and differentially expressed.The full length of the cDNA sequence of MVK gene was obtained by homologous cloning using the cDNA of S.baumii as a template and named as SbMVK(GenBank number MN793978).Sequence analysis showed that the full length of SbMVK gene cDNA sequence was 1041 bp,encoding 346 amino acid.The molecular weight of the SbMVK protein was predicted to be 36.6 ku,which belonged to GHMP kinase superfamily.Sequence alignment of amino acids revealed that SbMVK protein had the highest identity with the MVK protein of Ganoderma sinense.Moreover,the results of qRT-PCR and determination of triterpenoids content showed that the transcript level of SbMVK gene and triterpenoids content changed dynamically at different developmental stages,and the variation trend was basically the same.The results indicated that SbMVK gene plays an important role in MVA pathway of S.baumii,which will be useful for further revealing the function of the SbMVK gene of S.baumii and obtaining the strain with high triterpenoids content.
作者
刘增才
孙婷婷
孙健
马依莎
邹莉
LIU Zengcai;SUN Tingting;SUN Jian;MA Yisha;ZOU Li(College of Forestry,Northeast Forestry University,Harbin 150040,China;College of Food Engineering,Harbin University,Harbin 150086,China)
出处
《吉林农业大学学报》
CAS
CSCD
北大核心
2023年第5期547-552,共6页
Journal of Jilin Agricultural University
基金
中央高校基本科研业务费专项(2572018AA34)。
关键词
暴马桑黄
甲羟戊酸激酶
三萜
生物信息学分析
差异表达分析
Sanghuangporus baumii
mevalonate kinase
triterpenoids
bioinformatics analysis
differential expression analysis
