广西医科大学学报2024，Vol.41Issue(5)：668-675,8.DOI:10.16190/j.cnki.45-1211/r.2024.05.005
穿心莲ApbHLH80基因克隆及表达分析
Cloning and expression analysis of ApbHLH80 gene in Andrographis paniculata
摘要
Abstract
Objective:To clone the transcription factor gene ApbHLH80 of Andrographis paniculata(A.panicu-lata)and analyze its bioinformatics and expression.Methods:Primers were designed according to the transcrip-tome database of A.paniculata,and the full-length sequence of ApbHLH80 was cloned by PCR amplification and other methods.The physical and chemical nature of the protein encoded by this gene was analyzed and predicted through bioinformatics online tools.Reverse transcription-quantitative PCR(RT-qPCR)was used to analyze the expression pattern of this gene under methyl jasmonate treatment.And the content of andrographolide was deter-mined by high-performance liquid chromatography(HPLC).Results:Bioinformatics analysis showed that the open reading frame length of the gene contained 1,095 bp and encoded 364 amino acids.The amino acid se-quence encoded by this gene without signal peptide and transmembrane structure had a molecular weight of ap-proximately 39.96 ku and a theoretical isoelectric point of 5.73,and it's a hydrophilic protein in the nucleus.This protein was closely related to many species,such as Perilla frutescens var.Frutescens,Salvia miltiorrhiza,Salvia splendens and Salvia hispanica,all of which had the bHLH_AtbHLH_like conserved domain.The results of RT-qPCR analysis showed that the gene expression of ApbHLH80 was the highest in leaves,and its expression pat-tern was closely related to andrographolide synthesis in response to the induction of methyl jasmonate.Conclu-sion:The successful cloning of ApbHLH80 gene may regulate the andrographolide synthesis,providing theoreti-cal support for further exploration of its potential regulatory effects.关键词
穿心莲/ApbHLH80基因/基因克隆/生物信息学/表达分析Key words
Andrographis paniculata/ApbHLH80gene/gene cloning/bioinformatics/expression analysis分类
医药卫生引用本文复制引用
张洪波,钟楚,闫志刚..穿心莲ApbHLH80基因克隆及表达分析[J].广西医科大学学报,2024,41(5):668-675,8.基金项目
国家自然科学基金资助项目(No.82260744) （No.82260744）
广西自然科学基金资助项目(No.GXNSFAA2022035524 （）
No.2021GXNSFBA220011) （）
广西药用资源保护与遗传改良重点实验室资助项目(No.KL2020ZZ05 （）
No.KL2020ZZ03) （）
