| 大鳞副泥鳅角蛋白18基因cDNA的全长克隆与序列分析 |
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| 引用本文： | 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(2):362-369. |
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| 作者姓名： | 刘士力 吴卫君 张爱菊 练青平 王雨辰 辛建美 胡廷尖 贾永义 |
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| 作者单位： | 1. 浙江省淡水水产研究所/农业部淡水渔业健康养殖重点实验室/浙江省淡水水产遗传育种重点实验室，浙江湖州 313001; 上海海洋大学 水产与生命学院，上海 201306;2. 浙江省庆元县水利局水产技术推广站,浙江 庆元,323800;3. 浙江省淡水水产研究所/农业部淡水渔业健康养殖重点实验室/浙江省淡水水产遗传育种重点实验室,浙江湖州,313001 |
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| 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)，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)，Public Technology Research Grant from Zhejiang Province(2010C32033，2012C22059) |
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| 摘 要： | 角蛋白18(K18)是角蛋白的一种,为中间纤维蛋白,是细胞骨架的必要组成部分。采用RACE法克隆了大鳞副泥鳅K18基因cD NA的全长序列,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,克隆得到的大鳞副泥鳅K18基因cD NA序列全长为1 718 bp,其中包含1个长1 296 bp开放阅读框,编码431个氨基酸,蛋白质相对分子量48.66 ku。同源性分析显示,大鳞副泥鳅K18与泥鳅的相似性最高为98%,与斑马鱼、金鱼、虹鳟、白斑狗鱼等物种K18也存在不同程度的相似性。系统进化分析表明,大鳞副泥鳅与其他物种的K18系统发育同源,并与泥鳅的关系最为密切。通过实时荧光定量PCR分析,K18基因在大鳞副泥鳅的肠、肌肉、胃、心脏、肝脏、精巢、卵巢、脾脏等8种组织中都有表达,其中,肠表达最高,心脏最低。研究旨在为大鳞副泥鳅的分子生物学研究提供参考资料,并为更好地了解大鳞副泥鳅细胞骨架结构系统提供依据。
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| 关 键 词： | 角蛋白18( K18) 大鳞副泥鳅 克隆 定量 表达 |
Full-length cDNA Cloning and Sequence Analysis of Keratin 18 Gene from Paramisgurnus dabryanus |
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| 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. |
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| Keywords: | keratin 18( K18) Paramisgurnus dabryanus clone quantitative expression |
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