黄连重金属ATP酶基因CcHMA3的克隆和表达分析OACSTPCD

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.