水产学报2012，Vol.36Issue(9)：1376-1385,10.DOI:10.3724/SP.J.1231.2012.27888
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 )
摘要
Abstract
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 of FSHR gene 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 403 Y in EL I (extracellular loop, EL I ) and three protein kinase C phosphorylation sites are 634S643s;632T in C-terminal domain. The areas played the important role in the function of FSHR 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 of FSHR 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 of FSHR gene and the breeding of yellow catfish.关键词
黄颡鱼/促卵泡激素受体/基因克隆/半定量mRNA表达Key words
Pelteobagrus fulvidraco/ follicle-stimulating hormone receptor(FSHR) / gene clone/ RT-PCR mRNA expression分类
生物科学引用本文复制引用
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):1376-1385,10.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) （BZ200709）
