人HSPA8基因生物信息学分析及亚细胞定位

doi:10.13560/j.cnki.biotech.bull.1985.2019-0472

生物技术通报 ›› 2019, Vol. 35 ›› Issue (11): 82-88.doi: 10.13560/j.cnki.biotech.bull.1985.2019-0472

人HSPA8基因生物信息学分析及亚细胞定位

吴小敏, 赵佳福, 倪锴, 朱晓峰, 许厚强

1. 贵州大学高原山地动物遗传育种与繁殖教育部重点实验室贵州省动物遗传育种与繁殖重点实验室,贵阳 550025;
2. 贵州大学动物科学学院,贵阳 550025

收稿日期:2019-05-30 出版日期:2019-11-26 发布日期:2019-11-19
作者简介:吴小敏,女,硕士研究生,研究方向：遗传育种与繁殖;E-mail：1655367688@qq.com
基金资助:
国家自然科学基金项目（31860242）,教育部中国与美大、欧洲地区教育科研合作项目（教财司函[2017]860号）,贵州大学培育项目（黔科合平台人才[2017]5788-20）

Bioinformatics Analysis and Subcellular Localization of Human HSPA8 Gene

WU Xiao-min, ZHAO Jia-fu, NI Kai, ZHU Xiao-feng, XU Hou-qiang

1. Key Laboratory of Animal Genetics,Breeding and Reproduction in the Plateau Mountains Region of Ministry of Education（Guizhou University）/Key Laboratory of Animal Genetics,Breeding and Reproduction,Guiyang 550025;
2. College of Animal Sciences,Guizhou University,Guiyang 550025

Received:2019-05-30 Published:2019-11-26 Online:2019-11-19

摘要/Abstract

摘要： 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pEGFP-C1-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1 941 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。

关键词: HSPA8, 基因克隆, 生物信息学, 亚细胞定位

Abstract: 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-293T 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 1 941 bp in length and the molecular formula was C₃₁₁₁H₄₉₉₈N₈₆₀O₉₉₄S₁₇. The relative molecular weight was 70.89 kD,encoding a total of 646 amino acids. The secondary structure prediction showed HSPA8 protein consisted of the α-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.

Key words: HSPA8, gene cloning, bioinformatics, subcellular localization

吴小敏, 赵佳福, 倪锴, 朱晓峰, 许厚强. 人HSPA8基因生物信息学分析及亚细胞定位[J]. 生物技术通报, 2019, 35(11): 82-88.

WU Xiao-min, ZHAO Jia-fu, NI Kai, ZHU Xiao-feng, XU Hou-qiang. Bioinformatics Analysis and Subcellular Localization of Human HSPA8 Gene[J]. Biotechnology Bulletin, 2019, 35(11): 82-88.

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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

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