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角蛋白18(K18)是角蛋白的一种,为中间纤维蛋白,是细胞骨架的组成部分。为了解泥鳅(Misgurnus anguillicaudatus)K18序列特征及其在不同组织中的表达,研究采用3’-和5’-RACE法克隆K18基因c DNA全长序列,并运用实时荧光定量PCR技术探讨其在不同组织中的表达。结果表明,泥鳅K18基因c DNA全长序列1 694 bp,其中包含开放阅读框1 299bp,编码432个氨基酸,其序列具有Ⅰ型角蛋白序列DGKVVS,以及非常相似序列DNA(R/K)LAADDF。相似度分析显示,泥鳅K18与大鳞副泥鳅(Paramisgurnus dabryanus)的相似度最高为98%,与其它物种K18也存在不同程度的相似度。系统进化分析表明,泥鳅与其他物种的K18系统发育同源,且同陆生脊椎动物和硬骨鱼类同源,并与大鳞副泥鳅的关系最为密切。通过定量分析,K18基因在泥鳅肠、肌肉、心脏、胃、肝脏、精巢、卵巢、脾脏等8种组织都有表达,其中,肠表达最高,心脏最低。统计学分析表明,K18基因的表达在不同组织间存在显著性差异。
Keratin 18 (K18), a type of keratin, is an intermediate fibrin that is part of the cytoskeleton. In order to understand the characteristics of K18 sequence of Misgurnus anguillicaudatus and its expression in different tissues, the full-length cDNA sequence of K18 gene was cloned by 3’- and 5’-RACE methods and its real-time PCR Expression in tissue. The results showed that the full length cDNA sequence of K18 gene of Loach K18 contained 1 299 bp, encoding 432 amino acids. Similarity analysis showed that the similarity between loach K18 and Paramisgurnus dabryanus was 98%, and there was some degree of similarity with other species K18. Phylogenetic analysis showed that the loach has homology with K18 phylogeny of other species and is homologous to terrestrial vertebrate and teleost fish and has the closest relationship with Paramisgurnus dabryanus. Through quantitative analysis, K18 gene was expressed in eight tissues of intestinal loach, muscle, heart, stomach, liver, testis, ovary and spleen, with the highest expression in the intestine and the lowest in the heart. Statistical analysis showed that K18 gene expression in different tissues there is a significant difference.