| 摘要: |
| 采用同源克隆和末端快速扩增（RACE）方法，首次获得全长为1 319bp的半滑舌鳎膜孕激素受体（mPRα）的cDNA序列；将推断的氨基酸序列与其他物种mPRα氨基酸序列进行多重比较分析，发现存在7个跨膜区域。使用MEGA 4.0临位相联法和ClustalX方法对mPRα的氨基酸序列进行聚类分析和序列相似度分析。结果表明，半滑舌鳎mPRα与漠斑牙鲆、大西洋绒须石首鱼以及青鳉等鱼类的mPRα聚为一支，亲缘关系较近，相似度较高，分别为94%、93% 和90%；而与高等哺乳动物人和牛相似性均为53%，亲缘关系较远。应用半定量RT-PCR技术分析了mPRα mRNA在性成熟雌性半滑舌鳎不同组织的表达，结果表明,mPR α mRNA组织表达具有广泛性，但表达量存在差异，在脑、肾、脾和卵巢等组织表达丰富，在肝、胃和肌肉组织表达较弱。 |
| 关键词: 半滑舌鳎 膜孕激素 受体基因 基因克隆 组织表达 |
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| 基金项目:国家鲆鲽类产业技术体系(CARS-50)、国家自然科学基金(31201982) 、公益性农业行业专项项目(200903005)和留学人员科技活动项目择优资助经费 |
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| Molecular cloning and tissue expression analysis of membrane progestin receptor alpha gene (mPRα) from half smooth tongue sole Cynoglossus semilaevis Günther |
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| Abstract: |
| Full-length cDNA encoding membrane progestin receptor alpha gene (mPRα) was firstly cloned from half smooth tongue sole Cynoglossus semilaevis Günther by homology cloning and RACE-PCR analysis. The length of complete cDNA sequence of mPRα gene was 1 319 bp. Sequence alignment of deduced amino acid of tongue sole mPRα and amino acid of other species showed that there were seven transmembrane domains. The rooted phylogenetic tree was constructed by the neighbor-joining method of MEGA 4.0, and the identity of tongue sole mPRα with other representative sequences was analyzed by ClustalX. The results indicated that the tongue sole mPRα was clustered together with mPRα of other fish. The identity was 94%, 93% and 90% when compared with Southern flounder Paralichthys lethostigma, Atlantic croaker Micropogonias undulates and Japanese medaka Oryzias latipes, respectively. In contrast, identity among the mPRα of tongue sole and that of human Homo sapiens and cattle Bos taurus was low, only 53%. A semi-quantitative RT-PCR was developed to measure mRNA expression levels of mPRα gene of female tongue sole. Tissue expression analysis showed that mPRα mRNA was expressed widely in tongue sole, although the expression level was not homogeneous. mPRα transcripts were highly abundant in brain, kidney, spleen and ovary, less abundant in liver, stomach and muscle. |
| Key words: Cynoglossus semilaevis Günther mPRα gene Gene cloning Tissue expression |
