首页 | 期刊简介 | 编委会 | 投稿指南 | 常用下载 | 联系我们 | 期刊订阅 | In English
引用本文:张玉喜,陈松林.大菱鲆(Scophthalmus maximus)MHCⅡA基因全长cDNA的克隆与组织表达分析.海洋与湖沼,2007,38(3):221-226.
【打印本页】   【HTML】   【下载PDF全文】   查看/发表评论  下载PDF阅读器  关闭
←前一篇|后一篇→ 过刊浏览    高级检索
本文已被:浏览 1569次   下载 1736 本文二维码信息
码上扫一扫!
分享到: 微信 更多
大菱鲆(Scophthalmus maximus)MHCⅡA基因全长cDNA的克隆与组织表达分析
张玉喜1,2, 陈松林2
1.中国海洋大学海洋生命学院 青岛266003;2.农业部海洋渔业资源可持续利用重点开放实验室 中国水产科学研究院黄海水产研究所 青岛266071
摘要:
采用同源克隆和末端快速扩增(RACE)方法,得到1131bp的大菱鲆MHCⅡA全长cDNA片段。该序列包括720bp的开放阅读框(0RF),75bp的5′末端非编码区(UTR)以及336bp的3′末端非编码区。利用CLASTAL W1.8软件进行MHCⅡA基因的氨基酸序列比对和同源性比较,结果表明,大菱鲆MHCⅡA与牙鲆MHCⅡA的相似性最高,为69.8%,与真鲷、鲈鱼和丽鱼的相似性次之,分别为65.5%、69.6%和61.4%,与人、鼠和鲨鱼MHCⅡA的相似性仅为19.8%、31.6%和26.9%。应用RT-PCR分析其组织表达发现MHCⅡA基因在正常大菱鲆组织中均表达,但表达量在各个组织中不同,其中在鳃、脾、头肾、心脏、小肠和皮肤中有较强的转录本,中等程度表达于肝和血,在性腺和肌肉中表达最弱。
关键词:  大菱鲆  MHC  克隆  组织特异性表达
DOI:
分类号:
基金项目:国家重点基础研究发展规划(973)项目,2004B117403号;青岛市科技将才计划项目资助,2005—2008
附件
FULL LENGTH cDNA CLONING AND TISSUE EXPRESSION OF MAJOR HISTOCOMPATIBILITY COMPLEX (MHC) II A FROM TURBOT SCOPHTHALMUS MAXIMUS
ZHANG Yu-Xi1,2, CHEN Song-Lin2
1.College of Marine Life Science, Ocean University of China, Qingdao, 266003;2.Key Laboratory for Sustainable Utiliza-tion of Marine Fisheries Resource, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao, 266071
Abstract:
The full-length cDNA encoding turbot MHC class II A was cloned from turbot using homology cloning and RACE PCR approach. The MHC class II A was 1131bp in length, including 75bp 5′ terminal UTR, 720bp encoding region and 336bp 3' terminal UTR with a AU motif (ATTTA) that associated with mRNA instability and three canonical polyadenylation signals (AATAAA), followed by an additional 26bp poly (A) tail. The 720bp ORF was found to code a protein with 239 amino acid residues, and its secondary structure was analyzed using PHDsec Program. One N-glycosylation site (N-X-S/T) was observed in alpha 1 domain region. Two protein kinase C phosphorylation sites (S/T-X-R/K) were identified in alpha 2 domain region and five casein kinase II phosphorylation sites (S/T-X-X-D/E) were identified in alpha 1 and alpha 2. Four N-myristoylation sites (G-E/D/R/K/H/P/F/Y/W-X-X-S/T/A/G/C/N-G/I/T) were found in alpha 1, alpha 2 and transmembrane region. In addition, signature (F/Y-X-C-X-V/A-X-H) of immunoglobulins and MHC proteins occurred in alpha 2 domain region (residues 182-188). Alignment of deduced amino acid sequences of the MHC class II A and those of other vertebrates demonstrated that they shared many protein features common in those of other fish and mammalians. Phylogenetic analysis using CLASTAL W software showed that the putative MHC II A and that of Japanese flounder were close to each other in the first sister group in similarity of 69.8%. For other animals, the similarity was 65.5%, 69.6%, 44.4%, 61.4%, 47.6%, 46.3%, 43.6%, 19.8%, 31.6% and 26.9% from red sea bream, striped sea bass, carp, cichlid, rainbow trout, Atlantic salmon, zebrafish, human, mouse and nurse shark, respectively. Semi-quantitative RT-PCR analysis demonstrated that MHC class II A was ubiquitously expressed in tissues, but the expression level was locally unique. The MHC II A transcript was detected at a high level in head kidney, spleen, intestine, gill and heart, moderate in liver and blood, and low in gonad and muscle.
Key words:  Turbot Scophthalmus maximus, Major histocompatibility complex (MHC), Cloning, mRNA expression
版权所有 海洋与湖沼 Oceanologia et Limnlolgia Sinica Copyright©2008 All Rights Reserved
主管单位:中国科协技术协会 主办单位:中国海洋湖沼学会
地址:青岛市海军路88号  邮编:266400  电话:0532-82898753  E-mail:ols@qdio.ac.cn
技术支持:北京勤云科技发展有限公司