| 摘要: |
| 于2000年3月提取中华绒螯蟹眼柄总RNA，以此为模板，根据日本对虾的神经肽Pej-SG-Ⅳ设计兼并引物，进行RT-PCR扩增，在适宜的反应条件下，获得-特异性的产物，将该片段克隆到载体中进行测序。结果表明，中华绒螯蟹特异性cDNA片段由213个碱基组成。在生物信息数据库中查找由该cDNA推定的氨基酸序列的相似序列，可以发现有大量甲壳动物的CHH家族神经肽与它相似。在所有相似序列中，甲壳动物的蜕皮抑制激素(MIH)与它的相似度最高，这一结果提示由中华绒螫蟹的眼柄特异性cDNA片段推定的氨基酸序列可能是它的MIH片段。 |
| 关键词: 中华绒螫蟹, 蜕皮抑制激素, cDNA克隆, 序列分析 |
| DOI： |
| 分类号: |
| 基金项目:中国科学院知识创新工程项目，ZKCX-211号；国家自然科学基金资助项目，40106018号；山东省重点资助项目011110104号 |
附件 |
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| MOLECULAR CLONING AND SEQUENCE ANALYSIS OF cDNA ENCODING PARTIAL PUTATIVE MOLT-INHIBITING HORMONE FROM THE CRAB ERIOCHEIR SINENSIS |
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WANG Zai-Zhao, XIANG Jian-Hai, CUI Zhao-Xia
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Institute of Oceanology，The Chinese Academy of Sciences
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| Abstract: |
| Total RNAs were extracted from eyestalks of the crab Eriocheir sinensis. Eyestalk complementary DNAs were obtained from these total RNAs by reverse transcription. Based on the amino acid sequence of Pej-SG-Ⅳ, from Penaeus japonicas, two degenerate primers were designed. Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) was performed by using these eyestalk cDNAs an d the degenerate primers. A specific cDNA was cloned under proper reaction conditions. The specific cDNA was subcloned into pUCm-T vector for sequencing. The sequencing results demonstrated that the specific cDNA from the crab E.sinensis consists of 213 base pairs. The alignment of the deduced amino acid sequence from eyestalk cDNA of the crab E.sinensis with neuropeptides of CHH family from various crustaceans demonstrates that it has the highest identity an d homology with most MIHs from other crustaceans. The highest and the second highest amino acid identity percentages with MIH from the Shrimp Metapenaeus ensis and P.japonicus are 70.8% and 70.4%, respectively. It suggests that the deduced amino acid sequence from eyestalk cDNA of the crab E.sinensis is a part of its MIH neuropeptide of E.sinensis. |
| Key words: Eriocheir sinensis, Molt-inhibiting hormone, Complementary DNA cloning, Sequence analysis |
