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中国农学通报

中国农学通报 ›› 2015, Vol. 31 ›› Issue (8): 57-63.doi: 10.11924/j.issn.1000-6850.casb14100092

所属专题： 生物技术

• 水产 渔业 • 上一篇    下一篇

马氏珠母贝CyPB基因的克隆与表达分析

田荣荣,闫 芳,杜晓东,焦 钰,黄荣莲,王庆恒,邓岳文   

  1. (广东海洋大学水产学院,广东湛江 524025）
  • 收稿日期:2014-10-22 修回日期:2014-12-03 接受日期:2014-12-03 出版日期:2015-04-07 发布日期:2015-04-07
  • 通讯作者: 杜晓东
  • 基金资助:
    国家自然科学基金“马氏珠母贝珍珠质形成相关microRNA的鉴定与功能研究”(41206141)；国家自然科学基金“基于珍珠贝基因组测序分析的珍珠形成矿化关键基因和蛋白质的研究”(31272635)；国家自然科学基金“马氏珠母贝生长性状QTLs精细定位与相关基因功能分析”(31372526)。

Molecular Characterization and Expression Analysis of CyPB Gene from Pinctada martensii

Tian Rongrong, Yan Fang, Du Xiaodong, Jiao Yu, Huang Ronglian, Wang Qingheng, Deng Yuewen   

  1. (Fisheries College, Guangdong Ocean University, Zhanjiang Guangdong 524025)
  • Received:2014-10-22 Revised:2014-12-03 Accepted:2014-12-03 Online:2015-04-07 Published:2015-04-07

摘要： 为了探究亲环蛋白B(CyPB)在马氏珠母贝中的作用机制，运用cDNA末端快速扩增(RACE)技术克隆得到马氏珠母贝CyPB基因(PmCyPB)cDNA的全长序列，并且应用实时荧光定量PCR技术对PmCyPB基因在马氏珠母贝不同组织中的表达进行检测。结果显示，PmCyPB基因序列全长1266 bp，其中开放阅读框(ORF)为678 bp，编码225个氨基酸，5′-UTR为62 bp，3′-UTR为526 bp。预测其相对分子量为24829.3，理论等电点5.77。多序列比对结果显示PmCyPB基因与其他物种的CyPB具有较高的保守性。SMART软件对PmCyPB进行蛋白质序列分析，发现它包含亲环蛋白家族典型的肽脯氨酰顺反异构酶的结构域。实时荧光定量PCR数据分析表明，该PmCyPB基因在马氏珠母贝闭壳肌、肝胰腺、血细胞、外套膜、足、性腺、鳃这7种组织中均有表达，在外套膜表达量最高，其次是鳃。结果可为进一步阐述PmCyPB在马氏珠母贝中的免疫防御机制提供一定的理论基础。

关键词: 花粉管伸长, 花粉管伸长

Abstract: In order to explore the action mechanism of cyclophilin B in Pinctada martensii, the full length of PmCyPB gene were obtained using rapid amplification technology of cDNA ends. And expression of PmCyPB in different tissues was tested by Real-time PCR. The results showed that the total length of CyPB gene was 1266 bp, including 678 bp of the open reading frame (ORF) which encoding 225 amino acids, a 5-'UTR of 62 bp and a 3'-UTR of 526 bp. The predicted molecular weight was 24829.3, and the isoelectric point was 5.77. Multiple sequence alignment showed that PmCyPB was highly conservative in CyPB gene with other species. By SMART software analysis, we found that it contained the typical domain of the CyPB family, namely peptidylcis-transisomerase. In addition, real-time PCR indicated that PmCyPB could be expressed in all of the detected tissues, including adductor muscle, hepatopancreas, haemocytes, mantle, foot, gill, gonads, with the highest expression in mantle and the second in gill. The results could provide certain theoretical basis for the further study of PmCyPB in the immune response in mollusk.