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Cloning and Homology Analysis of HSP90 Gene of Carassius auratus by Consensus Degenerate Hybrid Oligonucleotide Primers
Authors:JIAO Xue  ZHANG Ya-bin  LU Yu-ting  ZU Xiu-jie  ZHANG Pei-jun  LI Yue-hong
Institution: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
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.
Keywords:Carassius auratus  degenerate primer  HSP90  
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