CODEHOP克隆鲫鱼HSP90基因及其同源性分析

doi:10.16431/j.cnki.1671-7236.2015.08.013

›› 2015, Vol. 42 ›› Issue (8): 2006-2010.doi: 10.16431/j.cnki.1671-7236.2015.08.013

CODEHOP克隆鲫鱼HSP90基因及其同源性分析

焦雪¹, 张雅斌², 卢玉婷¹, 祖岫杰², 张培军³, 李月红¹

1. 吉林农业大学动物科学学院, 长春 130118;
2. 吉林省水产科学研究院, 长春 130000;
3. 吉林省卫生监测检验中心, 长春 130000

收稿日期:2015-02-03 出版日期:2015-08-20 发布日期:2015-08-27
通讯作者: 李月红 E-mail:liyhong@sina.com
作者简介:焦雪(1992-),女,吉林长春人,硕士,研究方向:动物疾病及免疫,E-mail:455261939@qq.com
基金资助:
国家自然科学基金(30972191);农业部948项目(2014Z34);吉林省教育厅"十二五"科学技术研究重点项目(201443)

Cloning and Homology Analysis of HSP90 Gene of Carassius auratus by Consensus Degenerate Hybrid Oligonucleotide Primers

JIAO Xue¹, ZHANG Ya-bin², LU Yu-ting¹, ZU Xiu-jie², ZHANG Pei-jun³, LI Yue-hong¹

1. College of Animal Science, Jilin Agricultural University, Changchun 130118, China;
2. Jilin Fisheries Research Institute, Changchun 130000, China;
3. Health Monitoring and Inspection Center of Jilin Province, Changchun 130000, China

Received:2015-02-03 Online:2015-08-20 Published:2015-08-27

摘要/Abstract

摘要： 本研究旨在分析热休克蛋白90(HSP90)基因的保守性,并为寻找新的HSP基因,探索HSP90的应用提供理论数据。利用CODEHOP设计简并引物,采用PCR方法克隆鲫鱼的HSP90序列,并将扩增的序列克隆到pMD18-T载体中构建克隆载体后进行测序,再将测序结果通过BLAST进行同源性比对并构建系统进化树。结果显示,鲫鱼与鲤鱼的同源性为99%;与斑马鱼的同源性为92%。本试验设计出的引物简并度较低,Tm值的修改较为灵活,产物特异性强,为研究鲫鱼的功能基因提供基础保障,并为热休克蛋白在进化上的保守性研究奠定理论基础。

关键词: 鲫鱼; 简并引物; HSP90

Abstract: The purpose of this study was to analyze the conservative of heat shock protein 90(HSP90) gene, and provide theoretical data for the new HSP gene by exploring the application of HSP90.Using CODEHOP to design the degenerate primers, cloning Carassius auratus HSP90 sequence by PCR, and then the sequence was cloned into pMD18-T vector to construct clone vector after sequencing.Then sequencing homology were compared by BLAST and phylogenetic tree was constructed.The results showed that the homology was 99% between Carassius auratus and Cyprinus carpio; And the homology was 92% between Carassius auratus and Danio rerio.The degeneracy of primers designed in this study was low, the modification of Tm value was more flexible, and the product had high specificity.The study provided basis guarantee for research of Carassius auratus function genes, and laid the theoretical foundation for the study of heat shock proteins conservative in evolution.

Key words: Carassius auratus; degenerate primer; HSP90

中图分类号:

S917

焦雪, 张雅斌, 卢玉婷, 祖岫杰, 张培军, 李月红. CODEHOP克隆鲫鱼HSP90基因及其同源性分析[J]. , 2015, 42(8): 2006-2010.

JIAO Xue, ZHANG Ya-bin, LU Yu-ting, ZU Xiu-jie, ZHANG Pei-jun, LI Yue-hong. Cloning and Homology Analysis of HSP90 Gene of Carassius auratus by Consensus Degenerate Hybrid Oligonucleotide Primers[J]. , 2015, 42(8): 2006-2010.

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CODEHOP克隆鲫鱼HSP90基因及其同源性分析

Cloning and Homology Analysis of HSP90 Gene of Carassius auratus by Consensus Degenerate Hybrid Oligonucleotide Primers

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