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溶藻弧菌HY9901外膜蛋白OmpH的基因克隆及其生物信息学分析

, PP. 116-122

Keywords: 溶藻弧菌,基因克隆,OmpH,基因,外膜蛋白,生物信息学分析

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Abstract:

根据已发表的溶藻弧菌(Vibrioalginolyticus)全基因组序列设计1对特异性引物,PCR扩增溶藻弧菌HY9901株外膜蛋白OmpH的全长基因。结果显示,该基因(GenBank登录号:JX855924)全长573bp,共编码190个氨基酸残基。根据推导的氨基酸序列预测其分子量约为21.1kD,等电点为9.02。SignalP4.0、TMHMMServer2.0和SoftBerry-Psite预测结果显示,OmpH不存在信号肽与跨膜区,含有1个N-糖基化位点,6个磷酸化位点,1个内质网靶信号位点以及3个微体C末端靶信号位点。利用MAGE5.0软件,以邻位相连法构建OmpH系统进化树,结果显示,溶藻弧菌OmpH与副溶血弧菌(Vibrioparahaemolyticus)、哈氏弧菌(Vibrioharveyi)等有较近亲缘关系。采用多种参数成功预测了OmpH可能的B细胞抗原优势表位,分别是第5-10、106-110、120-125、158-163和175-180区段。利用SWISS-MODEL软件,模拟了OmpH亚基三维结构模型,且与其同源蛋白大肠杆菌(Escherichiacoli)Skp蛋白有相似构型。

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