文章摘要
刘淼,温海深,何峰,李吉方.黄颡鱼促卵泡激素受体基因的克隆及其在雌鱼生殖周期中的表达[J].水产学报,2012,36(9):1376~1385
黄颡鱼促卵泡激素受体基因的克隆及其在雌鱼生殖周期中的表达
Molecular cloning of follicle stimulating hormone receptor(FSHR)and expression analysis during the reproductive cycle in female yellow catfish(Pelteobagrus fulvidraco)
投稿时间:2011-12-21  修订日期:2012-06-13
DOI:DOI:10.3724/SP.J.1231.2012.27888
中文关键词: 黄颡鱼  促卵泡激素受体  基因克隆  半定量mRNA表达
英文关键词: Pelteobagrus fulvidraco  follicle-stimulating hormone receptor(FSHR)  gene clone  RT-PCR mRNA expression
基金项目:农业部水生动物遗传育种和养殖生物学重点开放实验室重点课题(BZ200709)
作者单位
刘淼 中国海洋大学水产学院 
温海深 中国海洋大学水产学院 
何峰 中国海洋大学水产学院 
李吉方 中国海洋大学水产学院 
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中文摘要:
      为研究黄颡鱼促卵泡激素受体(FSHR)基因的克隆以及该基因在卵巢发育周期中的表达情况,实验首先采用SmartTMRace 技术,获得了黄颡鱼促卵泡激素受体基因cDNA 全长序列,该基因全长2 340 bp,编码661aa,具有G蛋白偶联受体超家族(G protein-coupled receptor,GPCR)的7个跨膜螺旋区(transmembrane domain)。经分析发现,得到的FSHR基因序列具有9个富含亮氨酸的重复性序列(LRRs);5个潜在的N-端糖基化位点;蛋白激酶C(protein kinase C,PKC)磷酸化位点(403Y)位于第一个包外环,C-末端(C-terminal domain)G蛋白偶联区域,存在3个蛋白激酶C磷酸化位点(634S,643S,632T)。这些区域可能与FSHR的功能有密切关系。RT-PCR发现,FSHR基因在卵巢和脑中大量表达,在其他组织中均存在少量或微量的表达;在繁殖周期中,卵巢FSHR基因表达从Ⅲ期~Ⅳ期处于较高水平,在Ⅵ期时下降。研究推测,卵巢中FSHR基因参与黄颡鱼卵母细胞成熟及卵黄蛋白的积累过程。
英文摘要:
      Follicle-stimulating hormone(FSH)controls gonadal function in mammalian and many non-mammalian vertebrates through the interaction with their receptors(FSHR).Molecular cloning of the yellow catfish FSH receptor is reported together with changes in the gene expression throughout a reproductive cycle.This work describes the molecular cloning of the cDNA encoding the follicle stimulating hormone receptor(FSHR)of yellow catfish(Pelteobagrus fulvidraco)by means of reverse transcriptase polymerase chain reaction(RT-PCR),degenerate primers PCR amplification and 5′ and 3′ rapid amplification of cDNA ends(RACE)analyses.We cloned the complete cDNA sequence ofFSHRgene and the length is 2 340 bp,encoding a protein of 661 amino acids.This gene belongs to G protein coupled receptor(GPCR)family,which contains the seven conserved transmembrane helix(TM helix)domains.We found that the amino acids sequence contains nine Leucine rich repeats(LRRs),five potential N linked glycosylation sites and that only one protein kinase C phosphorylation site is 403Y in ELⅠ(extracellular loop,ELⅠ)and three protein kinase C phosphorylation sites are 634S,643S,632T in C-terminal domain.The areas played the important role in the function ofFSHR gene.The tissue distribution pattern showed high expression of FSHR gene in ovary and brain,and trace amount was detected weakly in other tissues.We also analyzed the mRNA expression level of FSHR gene in ovary at different stages of reproductive cycle.Result showed that the transcript level ofFSHR gene was high from Ⅲ to Ⅳ phases then followed by a substantial decline.These changes in expression profiles may imply differences in the functionality of the enzyme gene between different stages,suggesting an important role of FSHR gene in yellow catfish development.We inferred that the gene of FSHR in ovary might also be involved in the processes which regulate ovulation maturation and ovulation,and preferentially promote vitellogenin accumulation.The result has provided an improtant theoretical basis for the function ofFSHR gene and the breeding of yellow catfish.
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