目的 获得并分析黄连（Coptis chinensis Franch.）中自然抗性相关巨噬细胞蛋白（Natural Resistance-Associated Macrophage Protein，NRAMP）的编码基因。方法 根据黄连基因组测序结果中的NRAMP基因序列设计特异性引物，经PCR扩增得到黄连NRAMP5编码序列，进行TA克隆、测序及生物信息学分析。结果 得到黄连NRAMP5 cDNA序列，全长1611bp，编码536个氨基酸，通过与Genbank上的其他蛋白序列比对，将其命名为CcNRAMP5；序列分析表明黄连NRAMP5基因编码的氨基酸序列包含一个典型的NRAMP结构域（PF01566），是一种拥有11个跨膜结构域，亚细胞定位于质膜处的膜蛋白，具有疏水性强的特点。二级结构中肽链构象主要包括α-螺旋与无规则卷曲两部分；三级结构模型具有葡萄球菌锰转运突变体（MntH）的晶体结构。通过系统进化关系分析推测其可能具有二价金属离子转运功能。结论 首次通过克隆得到黄连NRAMP5基因，并运用生物信息学方法对其理化性质、结构特征等进行了分析预测，这些结果将为黄连NRAMP5基因的功能研究以及其对镉转运的调控机制提供理论依据。

Objective To obtain and analyze the gene encoding the natural resistance-associated macrophage protein NRAMP in Huanglian (Coptis chinensis Franch.).Methods Specific primers were designed based on the NRAMP gene sequence from the sequencing results of the genome of Coptis chinensis, and the coding sequence of NRAMP5 of Coptis chinensis was obtained by PCR amplification, TA cloning, sequencing and bioinformatics analysis.Results The sequence of NRAMP5 cDNA of Coptis chinensis, 1611bp in length, encoding 536 amino acids, was obtained and named as CcNRAMP5 by comparing with other protein sequences on Genbank; sequence analysis showed that the amino acid sequence encoded by Coptis chinensis NRAMP5 gene contains a typical NRAMP structural domain (PF01566), which was a 11 transmembrane structural domains, a membrane protein with subcellular localization at the plasma membrane. The secondary structure of the peptide chain conformation mainly consisted of two parts: α-helix and irregular coiled coil; the tertiary structure model had the crystal structure of the staphylococcal manganese transporter mutant MntH. In phylogenetic relationship CcNRAMP5 was closest in affinity to the borrelia NRAMP5 gene and poppy NRAMP5 gene, which may had the function of metal transporter.Conclusion The NRAMP5 gene of Coptis chinensis is cloned for the first time, and its physicochemical properties and structural characteristics are analyzed and predicted using bioinformatics methods. These results will provide a theoretical basis for the functional study of the NRAMP5 gene of Coptis chinensis and its regulatory mechanism on cadmium transport.

国家自然科学基金委员会面上项目（81973429）：基于全基因组解析NRAMP家族调控黄连镉富集分子机制，负责人：汪波； 国家自然科学基金委员会面上项目（32100299）：基于重测序的黄连种质资源综合评价及重要产量品质性状的全基因组关联分析，负责人：刘迪。