【作者】 吴松； 唐蕾； 傅明骏； 黄建华； 江世贵；

【Author】 WU Song;TANG Lei;FU Mingjun;HUANG Jianhua;JIANG Shigui;South China Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences;College of Aqua-life Science and Technology,Shanghai Fisheries University;

【机构】 中国水产科学研究院南海水产研究所； 上海海洋大学水产与生命学院；

【摘要】 在实验室已有的斑节对虾(Penaeus monodon)肝胰腺转录组数据中筛选出泛素基因(ubiquitin,Pm Ub)片段,通过RACE-PCR获得了Pm Ub基因的c DNA序列全长,利用生物信息学方法分析了Pm Ub的结构;利用实时定量PCR检验了Pm Ub分别在斑节对虾各组织及卵巢发育Ⅰ、Ⅱ、Ⅲ、Ⅳ和Ⅴ期的表达情况。结果显示,Pm Ub c DNA序列全长分别390bp,编码129个氨基酸,分子量约14.7k Da,等电点9.76;它第1-72个氨基酸为高度保守的泛素结构域,存在4个磷酸化位点;预测的Pm Ub二级结构由1个α螺旋6个β折叠构成。经BLAST同源性分析表明,该基因编码蛋白与凡纳滨对虾(Solenopsis invicta)、中华绒螯蟹(Megachile rotundata)等多种节肢动物Ub有很高的同源性。实时定量PCR结果表明,Pm Ub在卵巢Ⅱ期表达量最高,在各组织表达中,在脑神经、胸神经、心脏、肝胰腺、肠、肌肉、血淋巴和卵巢中均有表达,其中肝胰腺表达量最高,肌肉、卵巢表达次之。结果表明Pm Ub在斑节对虾卵巢发育过程中可能具有重要的作用。更多还原

【Abstract】 In the biological cell, the ubiquitin(Ub) proteasome pathway(UPP) is the major protein degradation pathways. It’s confirmed that Ub play an important role in the cell cycle and gametogeny. In this study, the P. monondon Ub gene(Pm Ub) was identified and characterized by using an approach which combines the data of transcriptome and rapid amplification of c DNA end(RACE). The expression profile of Pm Ub in different development stages of the ovary was determined by real-time quantitative PCR. The full length c DNA of the Pm-Ub gene is of 390 bp, which encodes a protein of 129 amino acids and whose estimated molecular mass is 14.7k Da and theoretical isoelectric point is 9.76. The encoding protein of Pm-Ub2 has 4 phosphorylation site, in which the 1-72 amino acids fall within the ubiquitin domain. The results of Blast and phylogenetic analyses suggest that Pm Ub is a new member of the Ub family. The results of real-time PCR show that expression was highest in the hepatopancreas, moderate in ovary and muscle, and very low in the brain, hemolymph, heart and intestine. Our results uggesting that the gene plays an important role in ovary development in Penaeus monodon更多还原