摘要
[目的]对黄芪bHLH转录因子进行克隆测序和生物信息学分析。[方法]根据黄芪转录组数据预测引物,以黄芪cDNA为模板,扩增获得黄芪bHLH转录因子片段,克隆测序后得到全长核苷酸序列,然后应用生信方法进行分析。[结果]结果表明来源于黄芪的bHLH转录因子全长855 bp,编码284个氨基酸,理论相对分子质量为71.76 kDa,理论等电点为5.18;亚细胞定位于细胞核,不存在跨膜区及信号肽,为非分泌蛋白;与其他物种同源基因的氨基酸序列分析表明黄芪bHLH与大豆的bHLH家族具有较高的同源性。[结论]黄芪bHLH转录因子基因的成功克隆和生物信息学分析为进一步阐明黄芪皂苷生物合成途径及其调控机制提供研究基础。
[Objective] Cloning,sequencing and bioinformatics analysis of bHLH transcription factor from Astragalus membranaceus.[Method] According to the transcriptome data of A.membranaceus,specific primers were designed.bHLH transcription factor fragment of A.membranaceus was amplified with cDNA as template,then the full-length nucleotide sequence was obtained after cloning and sequencing,and also analyzed by bioinformatics analysis.[Result] The total length of bHLH transcription factor from A.membranaceus was 855 bp,encoding 284 amino acids,its theoretical molecular weight was 71.76 kDa and the theoretical isoelectric point was 5.18.Subcellular localization was in cell nucleus,and there was no transmembrane region and signal peptide.Amino acid sequence analysis of homologous genes from other species showed that A.membranaceus originated bHLH had higher homology with G.max originated bHLH family.[Conclusion] The successful cloning and bioinformatics analysis of A.membranaceus originated bHLH provide a research basis for further elucidating the biosynthesis pathway and regulatory mechanism of astragalosides.
作者
杨李阳
YANG Li-yang(School of Basic Medical Sciences,Shanxi University of Chinese Medicine,Jinzhong 030619,China)
出处
《生物技术》
CAS
2023年第2期164-168,共5页
Biotechnology
基金
山西省回国留学人员科研项目(2020-132)。
关键词
黄芪
转录组测序
bHLH转录因子
基因克隆
生信分析
Astragalus membranaceus
transcriptome sequencing
bHLH transcription factor
gene cloning
bioinformatics analysis
