生物学杂志

生物学杂志

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三疣梭子蟹fps克隆及其在卵巢发育中的表达分析

  

  1. 宁波大学 海洋学院， 宁波 315211
  • 出版日期:2016-06-18 发布日期:2016-06-18
  • 通讯作者: 朱冬发，教授，研究方向为甲壳动物发育生物学，E-mail: zhudongfa@nbu.edu.cn
  • 作者简介:朱韬，专业方向为甲壳动物发育生物学
  • 基金资助:
    国家自然科学基金项目(41376152)；浙江省自然科学基金项目( LY13C190006)资助

Cloning of farnesyl diphosphate synthase (fps) from Portunus trituberculatus and its expression during the ovarian development

  1. School of Marine Sciences， Ningbo University， Ningbo 315211， China
  • Online:2016-06-18 Published:2016-06-18

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摘要： 法尼基焦磷酸合成酶(FPS)是类异戊二烯生物合成途径中的关键酶，参与甲壳动物甲基法尼酯(MF)等萜类激素的合成。为研究FPS在甲壳动物卵巢发育中的调控作用，采用RT-PCR和cDNA末端快速扩增技术(RACE技术)克隆得到了三疣梭子蟹fps(Ptfps)的序列全长(GenBank登录号：KM013803)。结果显示：Ptfps cDNA全长为2 360 bp，开放阅读框长1 290 bp，编码429个氨基酸。生物信息学分析发现PtFPS具有7个异戊烯基转移酶的保守结构域，与已公布的内华达古白蚁FPS同源性最高(达到58%)。实时荧光定量PCR分析结果显示：fps主要在大颚器(MO)中表达，远高于其它组织的表达水平 (P<0.05)，之后略有下降；在第2次卵巢发育过程中，fps的表达水平在Ⅲ期显著升高(P<0.05)，并在IV期达到最大。以上结果表明fps可能参与三疣梭子蟹卵巢发育的调控。

关键词: 三疣梭子蟹, 法尼基焦磷酸合成酶, 克隆, 卵巢发育, 表达水平

Abstract: Farnesyl diphosphate synthase (FPS) is a key enzyme in isoprenoid biosynthesis, participates in the synthesis of methyl farnesoate (MF) in the crustaceans. To study the regulatory role of FPS during ovarian development of the crustaceans, a full-length of fps (Ptfps, GeneBank accession number: KM013803) was cloned from Portunus trituberculatus by RT-PCR and rapid amplification of cDNA ends (RACE). The full-length of Ptfps was 2 360 bp and it contained an open reading frame (ORF) of 1 290 bp encoding a protein of 429 amino acid residues. Bioinformatics analysis found that PtFPS has seven prenyltransferase conserved regions, and exhibits the highest identity (58%) with FPS of Zootermopsis nevadensis. Quantitative real-time PCR analysis results showed that fps was mainly expressed in mandibular organ (MO), and the expression level was much higher than that in other tissues (P < 0.01). During the first ovarian development, the expression of fps significantly increased to the highest levels at stage Ⅳ (P < 0.05), and then slightly declined; During the second ovarian development, the expression of fps significantly increased to the highest level at stage Ⅲ (P < 0.05), and reached to the maximum at stage Ⅳ. The above results indicated that fps may closely related to the regulation of ovarian development in P. trituberculatus.

Key words: Portunus trituberculatus, farnesyl diphosphate synthase (FPS), gene cloning, ovarian development, expression