| 摘要: |
| 采用同源克隆和末端快速扩增(RACE)方法, 克隆了半滑舌鳎中 GnRHR 全长 cDNA。通过基因全长以及推断的氨基酸序列分析, 得知该序列包含一个 7tm-1 保守结构域, 为 G 蛋白偶联受体家族成员。利用 PHYLIP 3.5c 邻位相连法以及 DNAstar 中的 CLUSTAL W 方法对相应的氨基酸序列进行了聚类分析和序列相似度分析。结果表明, 半滑舌鳎 GnRHR 鳉 与鲈鱼、琥珀鱼、虹鳟以及青 等鱼类中的 GnRHR聚为一支, 亲缘关系较近, 且相似度较高, 分别为 90.1%、 89.7%、 79.0%以及 78.3%, 而与高等哺乳动物人、小鼠相似性仅分别为 18.8%和 16.2%, 亲缘关系较远。应用半定量 RT-PCR 技术分析其组织表达, 发现 GnRHR 广泛表达于各个组织, 但表达量差异较大, 在性腺、脑和肾中表达量较高, 其它组织较弱。 |
| 关键词: 半滑舌鳎, GnRHR, 克隆, 组织特异性表达 |
| DOI：10.11693/hyhz201104012012 |
| 分类号: |
| 基金项目:中央级公益性科研院所基本科研业务费专项资金, 2060302 号 |
附件 |
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| FULL LENGTH cDNA CLONING AND TISSUE EXPRESSION OF GONADOTRPIN-RELEASING HORMONE RECEPTOR (GnRHR) OF HALF SMOOTH TONGUE SOLE CYNOGLOSSUS SEMILAEVIS |
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LI Feng-Ling, WANG Qing-Yin, LI Zhao-Xin, ZHAI Yu-Xiu, GUO Meng-Meng
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Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences
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| Abstract: |
| Full-length cDNA encoding GnRHR was cloned from half smooth tongue sole Cynoglossus semilaevis by homology cloning and RACE PCR approach. According to the full length of the cDNA and deduced amino acid sequence, a 7tm-1 conservative domain was found and the cDNA was proved to be one of the G protein-coupled transmembrane family members. The rooted phylogenetic tree, analysis of identity of GnRHR, and other representative sequences were constructed by the neighbor-joining method within the package PHYLIP 3.5c and CLUSTAL W method, respectively. The results indicated that the GnRHR of the tongue sole was clustered together with those of all other fish GnRHR, for example, the similarity was 90.1%, 89.7%, 79%, 78.3% when compared with striped sea bass, amberjack, trout, and medaka, respectively. In contrast, similarity between the GnRHR of the tongue sole and those of human and mouse was much lower
(18.8% and 16.2%, respectively). Semi-quantitative RT-PCR analysis demonstrated that GnRHR was ubiquitously expressed in all tissues, although the expression level was not homogeneous. High levels of GnRHR transcript were detected in gonad, brain and kidney, and other tissues had lower levels. |
| Key words: Cynoglossus semilaevis, GnRHR, Cloning, Tissue-specific expression |
