吴松, 唐蕾, 傅明骏, 杨丽诗, 黄建华, 周发林, 江世贵. 斑节对虾泛素融合蛋白UbS27基因克隆与表达分析[J]. 南方水产科学, 2016, 12(1): 77-84. DOI: 10.3969/j.issn.2095-0780.2016.01.011
引用本文: 吴松, 唐蕾, 傅明骏, 杨丽诗, 黄建华, 周发林, 江世贵. 斑节对虾泛素融合蛋白UbS27基因克隆与表达分析[J]. 南方水产科学, 2016, 12(1): 77-84. DOI: 10.3969/j.issn.2095-0780.2016.01.011
shu
WU Song, TANG Lei, FU Mingjun, YANG Lishi, HUANG Jianhua, ZHOU Falin, JIANG Shigui. Cloning and expression analysis of PmUbS27 gene of black tiger shrimp (Penaeus monodon)[J]. South China Fisheries Science, 2016, 12(1): 77-84. DOI: 10.3969/j.issn.2095-0780.2016.01.011
Citation: WU Song, TANG Lei, FU Mingjun, YANG Lishi, HUANG Jianhua, ZHOU Falin, JIANG Shigui. Cloning and expression analysis of PmUbS27 gene of black tiger shrimp (Penaeus monodon)[J]. South China Fisheries Science, 2016, 12(1): 77-84. DOI: 10.3969/j.issn.2095-0780.2016.01.011
shu

斑节对虾泛素融合蛋白UbS27基因克隆与表达分析

Cloning and expression analysis of PmUbS27 gene of black tiger shrimp (Penaeus monodon)

    摘要
  • 摘要: 从构建的斑节对虾（Penaeus monodon）肝胰腺转录组数据中筛选出泛素核糖体融合蛋白UbS27基因（ubiquitin/ribosomal S27 fusion protein，PmUbS27）片段，利用SMART-RACE技术克隆出PmUbS27基因cDNA全长，并利用软件对其结构进行分析；利用实时荧光定量技术检测PmUbS27基因在斑节对虾不同组织及卵巢不同发育期的表达情况。结果显示，PmUbS27基因cDNA全长为514 bp，开放阅读框465 bp，编码154个氨基酸，含有泛素（1~72 aa）和核糖体蛋白（101~147 aa）2个结构域。以DNA为模板克隆得到的基因序列由3个内含子和4个外显子组成。实时荧光定量结果显示，PmUbS27基因在斑节对虾各组织中均有表达，但是在卵巢中表达量最高，其次为肝胰腺、血淋巴和精巢。PmUbS27在卵巢发育前期（Ⅱ期）和成熟期（Ⅴ期）的表达量显著高于其他各期（P0.05）。结果表明，PmUbS27参与斑节对虾卵巢发育的过程并可能在卵巢发育进程中起到重要的调控作用。

     

    Abstract: We screened the ubiquitin/ribosomal S27 fusion protein (PmUbS27) from hepatopancreas transcriptome data of Penaeus monodon and cloned the full-length PmUbS27 cDNA sequence by SMART-RACE technology. Besides, we studied the expression of ovaries of PmUbS27 gene at different developmental stages and in different tissues by real-time quantitative detection. The PmUbS27 cDNA was 514 base pairs (bp) long, containing a 3-UTR of 39 nucleotides and a 465 bp open reading frame (ORF) encoding a 154 amino acid polypeptide. The PmUbS27 protein contained the conserved Ub domain and ribosome structure. The DNA-cloned gene sequence consisted of four exons and three introns. PmUbS27 mRNA was ubiquitously detected in all tested tissues, comparatively high in the ovary, hepatopancreas and lymph. The expression level of PmUbS27 mRNA was significantly higher in ovary at Stage Ⅱ and Stage Ⅴ (P0.05). It is showed that PmUbS27 had an important role in oogenesis of P.monodon.

     

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