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在已构建的蜡梅花转录组数据库EST序列分析的基础上,采用RACE技术克隆获得蜡梅热激蛋白HSP70的cDNA序列,命名为CpHSP70-1(GenBank登录号:KR071130).该序列全长2 520bp,包括1 962bp的完整开放阅读框,编码653个氨基酸蛋白序列,含有3个HSP70家族典型基序.CpHSP70-1蛋白与其他真核生物的HSP70蛋白具有较高的同源性,聚类分析显示其属于定位于细胞核/细胞质的蛋白.亚细胞定位结果支持该蛋白分布于细胞核的预测.利用实时荧光定量PCR对CpHSP70-1基因的表达特性进行分析,其在各组织中均有不同程度的表达,其中在成熟叶片中表达量最高;在不同花发育阶段呈现持续稳定的表达模式;该基因在低温和高温诱导下表达量提高,而且对高温的响应更为敏感.
Based on the EST sequence analysis of the constructed C. przewalskii transcriptome database, the cDNA sequence of the HSP70 was cloned by RACE technology and named CpHSP70-1 (GenBank accession number: KR071130) , Including a complete open reading frame of 1 962bp, encoding a protein sequence of 653 amino acids, containing three typical motifs of HSP70 family.CpHSP70-1 protein has high homology with other eukaryotic HSP70 proteins, and cluster analysis Which belongs to the protein located in the nucleus / cytoplasm.The results of subcellular localization support the prediction of the protein distribution in the nucleus.The expression characteristics of CpHSP70-1 gene were analyzed by real-time fluorescence quantitative PCR and expressed in different degrees in all tissues , Which showed the highest expression level in mature leaves. The expression pattern was stable at different stages of flower development. The expression of this gene was enhanced under low temperature and high temperature induction and more sensitive to high temperature.