鲤两种孕激素受体基因克隆、表达及比较分析

doi:10.13560/j.cnki.biotech.bull.1985.2018-1089

摘要/Abstract

摘要： 孕激素受体（Progesterone receptor,Pgr）是介导孕激素调控鱼类性腺发育和生殖细胞成熟的受体。为研究鲤（Cyprinus carpio）2种Pgr时空表达模式和表达分化趋势,采用cDNA末端快速扩增方法克隆两种Pgr基因（Pgr1和Pgr2）。Pgr1和Pgr2的全长cDNA序列分别为2 713 bp和2 730 bp,均编码628个氨基酸。二者核酸和蛋白质一致性分别为91%和90%。这两个基因都含有锌指结构域和核激素受体域同源超家族两个功能域。进化分析将鲤Pgr1和鲫Pgr1聚在一起,鲤Pgr2和鲫Pgr2聚为一支;这两支再与斑马鱼Pgr汇成一支。鲤Pgr1和Pgr2的非同义替换率与同义替换率比值为0.360,表明Pgr1和Pgr2受负选择压力。实时荧光qPCR分析表明,Pgr1和Pgr2在性腺的表达显著高于其他组织,说明二者可能参与性腺发育过程。催产素诱导后,Pgr1和Pgr2在精液和卵细胞的表达量高于受精卵,表明这两种基因与鲤生殖细胞成熟相关。大多数状态下Pgr1表达量显著高于Pgr2,表明Pgr1是介导鲤性腺发育和生殖细胞成熟的主表达基因。

关键词: 孕激素受体, 鲤, 实时荧光定量PCR, 性腺, 生殖细胞

Abstract: Progesterone receptor（Pgr）is a receptor mediating the regulation of progesterone on fish gonad development and germ cell maturation. To study the expression pattern and differential expression tendency of two common carp（Cyprinus carpio）Pgr genes,Pgr1 and Pgr2 were cloned using rapid amplification of cDNA ends. The full-length cDNA sequence lengths of Pgr1 and Pgr2 were 2 713 bp and 2 730 bp,both of which encoded proteins of 628 amino acids. They had a nucleotide identity of 91% and a protein identity of 90%. They both contained two domains,zinc finger domain and nuclear hormone receptor domain homologous superfamily. In the phylogenetic analysis,common carp Pgr1 and goldfish Pgr1 were clustered together and common carp Pgr2 and goldfish Pgr2 were grouped together. These two branches were then clustered with zebrafish Pgr. The ratio of the number of non-synonymous substitutions per non-synonymous site to the number of synonymous substitutions per synonymous site between Pgr1 and Pgr2 was 0.360,suggesting that they were under negative selection. qRT-PCR analysis demonstrated that Pgr1 and Pgr2 had higher expression levels in the gonads than in the other tissues,revealing that both genes might be associated with the gonad development of common carp. They had higher expression levels in sperms and oocytes than in embryos after induced by spawning drugs,indicating that they might be involved in germ cell maturation. In the most examined cases,significantly higher Pgr1 expression level was revealed compared with Pgr2,suggesting that Pgr1 plays the dominate role in common carp gonad development and germ cell maturation.

Key words: progesterone receptor（Pgr）, common carp, qRT-PCR, gonad, germ cell

朱锐, 叶雨情, 王雅欣, 杨晨茹, 王红伟, 孙晓晴, 张研, 李尚琪, 李炯棠. 鲤两种孕激素受体基因克隆、表达及比较分析[J]. 生物技术通报, 2019, 35(7): 46-53.

ZHU Rui, YE Yu-qing, WANG Ya-xin, YANG Chen-ru, WANG Hong-wei, SUN Xiao-qing, ZHANG Yan, LI Shang-qi, LI Jiong-tang. Cloning,Expression and Comparative Analysis of Two Progesterone Receptor Genes in Cyprinus carpio[J]. Biotechnology Bulletin, 2019, 35(7): 46-53.

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