江西农业大学学报2016，Vol.38Issue(2)：362-369,8.DOI:10.13836/j.jjau.2016051
大鳞副泥鳅角蛋白18基因cDNA的全长克隆与序列分析
Full-length cDNA Cloning and Sequence Analysis of Keratin 18 Gene from Paramisgurnus dabryanus
摘要
Abstract
Keratin 18 ( K18) is one of the keratins in the intermediate filament protein family,and is an integral part of the cytoskeleton.The full⁃length cDNA sequence of the K18 gene was cloned from Paramisgur⁃nus dabryanus using RACE, and its expression in different tissues was explored using real⁃time quantitative PCR assays.This novel study showed that the cloned cDNA full⁃length sequence of K18 was 1 718 bp long,and contained a 1 296 bp open reading frame that encoded 431 amino acids. The molecular weight of the protein was 48.66 ku.Homology analysis showed that Paramisgurnus dabryanus had the highest similarity of K18 with Misgurnus anguillicaudatus ( up to 98%);lower levels of similarity were found with other species,including ze⁃bra fish ( Danio rerio) ,goldfish ( Carassius auratus) ,rainbow trout ( Oncorhynchus mykiss) ,and northern pike (Esox lucius).Phylogenetic analysis confirmed that the K18 gene of Paramisgurnus dabryanus was phylogeneti⁃cally homologous with other species, and had closest relationship with Misgurnus anguillicaudatus. The K18 gene was found to be expressed in the intestine,muscle,stomach,heart,liver,testis,ovary,spleen eight tissue types within Paramisgurnus dabryanus.The highest expression level of K18 according to RT⁃qPCR was found in the intestine,and the lowest was in the heart.This study was designed to provide a reference database regarding K18 in Paramisgurnus dabryanus,and to provide the basis for the better understanding of its cytoskeletal sys⁃tem in future research.关键词
角蛋白18( K18)/大鳞副泥鳅/克隆/定量/表达Key words
keratin 18( K18)/Paramisgurnus dabryanus/clone/quantitative/expression分类
农业科技引用本文复制引用
刘士力,吴卫君,张爱菊,练青平,王雨辰,辛建美,胡廷尖,贾永义..大鳞副泥鳅角蛋白18基因cDNA的全长克隆与序列分析[J].江西农业大学学报,2016,38(2):362-369,8.基金项目
浙江省淡水水产遗传育种重点实验室建设项目（2014F10037）、浙江省重大科技专项（2013C02009）和浙江省公益技术研究项目（2010C32033，2012C22059） Project supported by Construction Project of the Key Laboratory of the Freshwater Aquatic Animal Genetic and Breeding of Zhejiang Province （2014F10037），the Grand Science and Technology Special of Zhejiang Province （2013C02009） and Public Technology Research Grant from Zhejiang Province （）
