马氏珠母贝热休克蛋白HSP60基因的克隆与表达分析Cloning and Express Characters of HSP60 gene from Pinctada martensii
王志新;梁海鹰;杜晓东;黄荣莲;邓岳文;王庆恒;焦钰;
摘要(Abstract):
运用cDNA末端快速扩增(RACE)技术,克隆马氏珠母贝(Pinctada martensii)热休克蛋白HSP60基因cDNA全序列。序列全长2495 bp,开放阅读框(ORF)1734 bp,编码577个氨基酸,预测的分子量约为61.79 ku,理论等电点为5.49。5'非翻译区(5'UTR)长150 bp,3'非翻译区(3'UTR)长611 bp。同源性比对分析结果,马氏珠母贝HSP60基因与与太平洋长牡蛎(Crassostrea gigas)和光滑双脐螺(Biomphalaria glabrata)的同源性较高,达到75%。氨基酸序列分析显示,马氏珠母贝HSP60氨基酸序列具有典型的mt-HSP60特征序列、C-末端典型的GGM重复基序和一个ATP结合结构域。荧光定量数据分析发现,该基因在闭壳肌、外套膜、血淋巴、肝胰腺、性腺、鳃等6个组织中具有表达,在肝胰腺中表达量最高,鳃中次之,而在血液和闭壳肌中仅有少量表达;在脂多糖刺激后,该基因表达水平上调,12 h后达到最大值,之后又逐渐下调,均高于对照组,差异具统计学意义(P<0.05)。
关键词(KeyWords): 马氏珠母贝;HSP60;基因克隆;表达分析
基金项目(Foundation): 国家自然科学基金(31272635);; 广东省科技计划项目(2012A031100010)
作者(Authors): 王志新;梁海鹰;杜晓东;黄荣莲;邓岳文;王庆恒;焦钰;
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