人HSPA8基因生物信息学分析及亚细胞定位

Bioinformatics Analysis and Subcellular Localization of Human HSPA8 Gene

探究人HSPA8基因生物信息学分析与亚细胞定位。利用用RT-PCR方法扩增HSPA8基因CDS区,利用生物信息学工具对其理化性质、二级结构、三级结构进行分析并构建其遗传进化树。运用酶切、连接等方法构建重组真核表达载体pEGFPC1-HSPA8,转染HEK-293T细胞,采用荧光共定位的方法观察其亚细胞定位。成功克隆人HSPA8基因CDS区,生物信息学分析结果显示:人HSPA8基因CDS区全长1941 bp,分子式为C3111H4998N860O994S17,相对分子量为70.89 kD,共编码646个氨基酸;二级结构预测显示HSPA8蛋白以α-螺旋(占41.64%)和无规则卷曲(32.20%)为主;HSPA8核苷酸同源性及遗传进化树分析表明人与黑猩猩亲缘关系最近。荧光共定位结果显示HSPA8蛋白均匀分布于整个细胞。HSPA8蛋白的生物信息学分析及亚细胞定位研究为进一步探究HSPA8基因胞内功能奠定基础。

This work is to investigate the bioinformatics analysis and subcellular localization of human HSPA8 gene.The HSPA8 gene CDS region was amplified by RT-PCR method,and its physicochemical properties,secondary structure and tertiary structure were analyzed by bioinformatics tools and its phylogenetic tree was constructed.The recombinant eukaryotic expression vector pEGFP-C1-HSPA8 was constructed by restriction enzyme digestion and ligation,and transfected into HEK-293 T cells.The subcellular localization was observed by fluorescence colocalization.The result showed that the CDS region of human HSPA8 gene was successfully cloned.The results of bioinformatics analysis showed that the human HSPA8 gene CDS region was 1941 bp in length and the molecular formula was C3111 H4998 N860 O994 S17.The relative molecular weight was 70.89 kD,encoding a total of 646 amino acids.The secondary structure prediction showed HSPA8 protein consisted of theα-helix(41.64%)and random coil(32.20%)dominantly.HSPA8 nucleotide homology and genetic phylogenetic tree analysis showed that humans and chimpanzees were closely related.Fluorescence colocalization results showed that HSPA8 protein was evenly distributed throughout the cell.The bioinformatics analysis and subcellular localization of HSPA8 protein laid the foundation for further exploring the intracellular function of HSPA8 gene.