文章摘要
陈晓燕,温海深,何峰,李吉方,陈彩芳,张葭人,金国雄,史宝,史丹,杨艳平.半滑舌鳎促滤泡激素受体基因克隆及其在雌鱼生殖周期中的表达[J].水产学报,2010,34(9):1309~1318
半滑舌鳎促滤泡激素受体基因克隆及其在雌鱼生殖周期中的表达
Cloning of FSHR and expression analysis during the reproductive cycle in female Cynoglossus semilaevis Günther
投稿时间:2010-04-03  修订日期:2010-07-15
DOI:10.3724/SP.J.1231.2010.06896
中文关键词: 半滑舌鳎  促滤泡激素受体  基因克隆  mRNA表达  雌二醇
英文关键词: Cynoglossus semilaevis Günther  follicle-stimulating hormone receptor(FSHR)  gene clone  mRNA expression  17β-Estradiol
基金项目:国家“十一五”科技支撑计划(2006BAD09A01)
作者单位E-mail
陈晓燕 中国海洋大学海水养殖教育部重点实验室  
温海深 中国海洋大学海水养殖教育部重点实验室 wenhaishen@ouc.edu.cn 
何峰 中国海洋大学海水养殖教育部重点实验室  
李吉方 中国海洋大学海水养殖教育部重点实验室  
陈彩芳 中国海洋大学海水养殖教育部重点实验室  
张葭人 中国海洋大学海水养殖教育部重点实验室  
金国雄 中国海洋大学海水养殖教育部重点实验室  
史宝 中国海洋大学海水养殖教育部重点实验室  
史丹 中国海洋大学海水养殖教育部重点实验室  
杨艳平 中国海洋大学海水养殖教育部重点实验室  
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中文摘要:
      通过简并引物扩增及RACE技术,首次克隆了半滑舌鳎促滤泡激素受体(FSHR)基因cDNA全长序列。该基因全长3 105 bp,编码704个氨基酸,含有典型的跨膜螺旋结构区域(TM helix),属于糖蛋白激素受体(GHR)家族。该基因编码区共有19个Ser,4个Thr和5个Tyr磷酸化位点;3个潜在的N基端糖基化位点,分别是62NISS,226NGIK和356NSTS;441T为潜在的PKC位点。基酸序列比对结果表明,半滑舌鳎FSHR含有GpHR家族的特殊信号序列,如82CCAF,481ERW,594FTD和667NPFLY,含有10个LRRs(LRR1-10)。组织表达分析表明,FSHR表达广泛,除在性腺中大量表达外,其他非性腺组织也有表达,尤其以脾、肾、头肾表达最较高。采用125Ⅰ标记放射免疫分析法(RIA)测定雌鱼血清中E2含量的季节变化,发现4月时E2含量最低,10月时含量最高。卵巢中FSHR mRNA在10月表达量最高,1月最低。
英文摘要:
      The complete cDNA sequence of follicle stimulating hormone receptor(FSHR)gene was cloned by degenerate primer PCR amplification and RACE cDNA amplification for the first time.The length of the cDNA is 3105 bp,and it encodes a protein of about 704 amino acids,which contains the conserved seven transmembrane helix domains(TM-helix),and it belongs to glycoprotein hormone receptor(GHR)family.Phosphorylation site predictions identified 19 Ser,4 Thr and 5 Tyr potential phosphorylation sites in tongue sole,and only one protein kinase C phosphorylation site(441T)was predicted.The Clustal X alignment also revealed the presence of specific signature sequences(e.g.82CCAF,481ERW,594FTD and 667NPFLY),which are highly conserved in GpHRs.And comparison with the halibut FSH-R also allowed the identification of 10 imperfect LRRs in the FSHR of Cynoglossus semilaevis.Tissue expression analysis showed that FSHR mRNA is expressed widely in C. semilaevis. Except the high levels in gonads,strong amplification signals were also detected in extragonadal tissues including the spleen,kidney and head kidney.Plasma 17β-Estradiol(E2)seasonal changes are detected by radio immunoassay(RIA).The result shows that the lowest level appears in April and the highest peak of a year is in October.And we analyzed the relative expression level of FSHR in reproductive cycle of female.There is the highest level in October and the lowest level in January.
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