| 摘要: |
| 采用RT-PCR、RACE等方法, 获得了拟穴青蟹PP2A调节亚基B(PP2A-B)的cDNA全序列, 全长2040bp, 开放阅读框(ORF)为1332bp, 可编码443个氨基酸残基。同源分析显示, 该基因编码的蛋白与其它一些物种具有很高相似性, 推测PP2A-B基因具有很高的保守性。经荧光定量PCR检测, PP2A-B基因在拟穴青蟹多个组织中有表达, 且在脑、卵巢、鳃中表达量较高。在拟穴青蟹卵巢发育过程中, PP2A-B 基因在卵巢未发育期(I期)表达量最高, 此后各期逐渐下降, 推测PP2A-B在卵巢中可能以PP2A全酶的形式存在, 抑制卵巢发育。 |
| 关键词: PP2A-B, 拟穴青蟹, 基因克隆, 荧光定量PCR, 卵巢发育 |
| DOI：10.11693/hyhz201205009009 |
| 分类号: |
| 基金项目:国家自然科学基金资助项目, 40406030 号, 41076081 号 |
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| CLONING AND EXPRESSION ANALYSIS OF THE REGULATORY SUBUNIT B GENE OF PP2A IN THE MUD CRAB SCYLLA PARAMAMOSAIN |
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LIU Xue-Liang, JIN Zhu-Xing, HUANG Hui-Yang, YE Hai-Hui, LI Shao-Jing
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College of Ocean and Earth Sciences, Xiamen University
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| Abstract: |
| In this paper, the regulatory subunit B geneof PP2A (PP2A-B) was isolated from the mud crab, Scylla paramamosain using RT-PCR and RACE methods. The obtained full-length cDNA of PP2A-B was 2040bp with an open reading frame of 1332bp encoding a putative peptide of 443 amino acid. By alignment, the amino acid sequence of S. paramamosain PP2A-B showed high homology with those of some other animals. It suggested PP2A-B was highly conservative. Real-time PCR showed that the PP2A-B gene was expressed in various tissues, and highly expressed in brain, ovary and gill. The PP2A-B mRNA expression during ovarian development indicated that the expression of PP2A-B was obviously high at undeveloped stage and decreased gradually from stage I (undeveloped stage) to stage V (ripe stage). We hypothesized that PP2A-B may exist in the form of PP2A holoenzyme in ovary and play a suppressive role on the ovarian development of the mud crab. |
| Key words: PP2A-B, Scylla paramamosain, Gene cloning, Real-time RT-PCR, Ovarian development |
