Cloning and Expression Analysis of Keratin 18 Gene in Loach（Misgurnus anguillicaudatus）

doi:10.13560/j.cnki.biotech.bull.1985.2016.05.015

Abstract

Abstract: Keratin 18（K18），one of the keratin，is the intermediate filament proteins and an integral part of the cytoskeleton. In order to understand the sequence characteristics and expression of K18，cDNA full-length sequence of K18 gene was cloned from loach（Misgurnus anguillicaudatus）using RACE，and its expression in different tissues was explored using real-time quantitative PCR. The results showed that the cDNA full-length of cloned K18 gene was 1 694 bp，containing a 1 299 bp open reading frame encoding 432 amino acids. There were the sequence DGKVVS，and the very similar sequence DNA（R/K）LAADDF of type I keratin. Identity analysis revealed that loach had the highest identity up to 98% with large-scale loach（Paramisgurnus dabryanus）in K18，and there were various levels of identity with other species. Phylogenetic analysis demonstrated that the K18 gene of loach was in phylogenetic homology with other species，and in true orthologs with terrestrial vertebrates（tetrapods）and teleosts，moreover，closest with large-scale loach. K18 gene was expressed in the 8 tissues of intestine，muscle，heart，stomach，liver，testis，ovary，spleen，the highest in the intestine，and the lowest in the heart. Statistical analysis showed that the differences of expression among different tissues were significant.

Key words: keratin 18（K18）, cDNA clone, loach（Misgurnus anguillicaudatus）, quantitative expression

WU Wei-jun,LIU Shi-li,LI Qian,WANG Yu-chen,LIAN Qing-ping,HU Ting-jian,JIA Yong-yi,JIANG Wen-ping,LI Xi-lian. Cloning and Expression Analysis of Keratin 18 Gene in Loach（Misgurnus anguillicaudatus）[J]. Biotechnology Bulletin, 2016, 32(5): 114-123.

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