低温冷害是影响玉米生产的重要环境因素之一。基于水稻中调控耐低温的OsCOLD1 基因编码序列，利用同源克隆方法，获得了玉米 ZmCOLD1基因，进而对ZmCOLD1基因的分子特征和编码蛋白的亚细胞定位进行分析，并进行ZmCOLD1基因过表达遗传转化载体的构建。结果表明，ZmCOLD1基因编码区长度为 1647bp，编码548个氨基酸，理论等电点( pI) 4.95，估计分子量137457.13Da，属于疏水蛋白，利用ClustalX进行蛋白质序列的比对， 进行ZmCOLD1基因系统进化树分析，ZmCOLD1氨基酸序列与水稻的亲缘关系较近。通过构建目的蛋白与 GFP 融合的表达载体pCAMBIA1302-GFP-ZmCOLD1并注射烟草，发现 ZmCOLD1蛋白主要集中在细胞质膜中。

Low temperature is one of primary environmental factors limiting maize production.In this study, based on the method of homology cloning, the OsCOLD1 sequence was used as a template to obtain the ZmCOLD1 gene.Then the molecular characteristics and subcellular localization of ZmCOLD1 gene were analyzed, and the genetic overexpression transformation vector of ZmCOLD1 was constructed.The results showed that the CDS length of ZmCOLD1 was 1647 bp, encoding 548 amino acids with a molecular weight of about 137457.13Da and a isoelectric points of 4.95, belonging to the hydrophobic protein. ClustalX was used to compare protein sequences and analyze the phylogenetic tree of ZmCOLD1 gene. ZmCOLD1 amino acid sequence was closely related to rice. By constructing the expression vector pCAMBIA1302-GFP-ZmCOLD1 fused with GFP and injecting tobacco, ZmCOLD1 protein was mainly concentrated in the cytoplasmic membrane.

吉林省主粮作物良种科技创新重大专项-玉米分子育种技术研究与应用(20210302003NC)