中药材2023，Vol.46Issue(12)：2969-2974,6.DOI:10.13863/j.issn1001-4454.2023.12.010
黄连重金属ATP酶基因CcHMA3的克隆和表达分析
Cloning and Expression Analysis of Heavy Metal ATPase Gene CcHMA3 in Coptis chinensis
摘要
Abstract
Objective:To clone the heavy metal ATPase gene CcHMA3 of Coptis chinensis and analyze its encoding protein bioinfor-matics and differential expression.Methods:Based on the genome and transcriptome data of Coptis chinensis,the full-length cDNA of CcHMA3 was amplified by PCR and bioinformatic analysis was performed.Real-time quantitative PCR(qRT-PCR)was used to detect the expression difference of CcHMA3 gene in Coptis chinensis.Results:CcHMA3 gene was cloned.The open reading frame length was 3 030 bp,encoding 1 009 amino acids.The relative molecular weight of the protein was 109.03 kD and the theoretical isoelectric point was 6.7.The protein sequence contained 6 transmembrane domains and 2 conserved domains.Subcellular localization prediction indica-ted that CcHMA3 was located on the plasma membrane and had certain hydrophilicity.The results of qRT-PCR showed that the relative expressions of CcHMA3 gene in different tissues of Coptis chinensis were from high to low as fibrous root,rhizome,petiole and leaf.After Cd treatment,the expression of CcHMA3 gene in fibrous roots was significantly higher than that in other tissues(P<0.05),and the ex-pression of CCHMA3 gene was significantly different in response to different Cd stress treatment times.Conclusion:This study lays an experimental foundation for further research on the regulatory mechanism of CcHMA3 gene in the heavy metal cadmium metabolism of Coptis chinensis,and further search for possible blocking methods.关键词
黄连/CcHMA3/基因克隆/镉胁迫/表达分析Key words
Coptis chinensis Franch./Cc HMA3/Gene cloning/Cadmium stress/Expression analysis分类
医药卫生引用本文复制引用
程华春,王文斌,莫静,李小芳,聂晶,汪波..黄连重金属ATP酶基因CcHMA3的克隆和表达分析[J].中药材,2023,46(12):2969-2974,6.基金项目
国家自然科学基金项目(81973429) （81973429）
中共湖北省委组织部青年拔尖人才培养计划项目(2021-10-18) （2021-10-18）
