斑节对虾泛素融合蛋白UbL40基因的克隆及表达分析

Molecular characterization and expression profiles of Ub S27 gene from black tiger shrimp(Penaeus monodon)

从斑节对虾（Penaeus monodon）肝胰腺转录组数据中筛选获得泛素融合蛋白Ub L40基因（ubiquitin/ribosomal L40 fusion protein,Pm Ub L40）的部分序列,利用RACE技术克隆其c DNA全长序列,通过生物信息学进行结构分析;利用实时定量PCR技术检测该基因在斑节对虾不同组织及卵巢不同发育期的表达情况。结果表明,Pm Ub L40基因c DNA全长571 bp,开放阅读框（Open reading frame,ORF）长390 bp,编码129个氨基酸,预测分子量约14.7 ku,理论等电点为9.76。预测氨基酸序列的N-末端（1～76aa）为高度保守的泛素结构域,C-末端（77～129aa）为典型的核糖体蛋白L40结构域。多重序列比对表明不同物种的Ub L40在进化过程中高度保守。实时定量PCR结果显示Pm Ub L40基因在肝胰腺的表达量最高,肌肉、卵巢次之,其他组织的表达较低;在卵巢发育过程中Pm Ub L40基因在II期表达最高,其次是V期,显著高于I、III、IV期。研究结果表明Pm Ub L40基因在斑节对虾卵巢发育过程中可能具有重要作用。

Penaeus monodon ubiquitin/ribosomal L40 fusion protein gene （ PmUbL40 ） was identified and characterized by using an approach which combines the data of transcriptome and rapid amplification of cDNA end （ RACE ） . The expression profile of PmUblAO in different development stages of the ovary and various tissues were determined by real-time quantitative PCR. The full length cDNA of PmUbL40 gene was of 390bp, which encoded a protein of 129 amino acids with estimated molecular mass of 14.7 ku and theoretical isoelectric point of 9.76. The encoded protein consists ubiquitin domain （ 1-76aa ） and ribosomal L40 domain （ 77-129aa ） . Real-time quantitative PCR results showed that PmUbL40 was expressed high in hepatopancreas than in other tissues. And PmUbL40 was expressed the highest in the ovary stay II, and than in ovary stage V, which indicated that PmUbL40 may play an important role in ovary development and oogenesis.