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溶藻弧菌附着定植因子ACFA原核载体构建、表达条件优化及多克隆抗体制备

DOI: 10.7668/hbnxb.2015.01.007, PP. 35-41

Keywords: 溶藻弧菌,ACFA蛋白,正交试验,多克隆抗体

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Abstract:

为构建水产致病菌溶藻弧菌附着定植因子ACFA原核表达载体,优化ACFA蛋白的诱导表达条件,制备ACFA蛋白小鼠多克隆抗体。通过分子克隆获得ACFA蛋白的表达菌株,利用切胶纯化获得ACFA蛋白,免疫小鼠制备多克隆抗体,采用ELISA法检测抗体滴度为1:3200倍,Western-Blotting法检测抗血清特异性较好。通过正交试验,获得ACFA菌株的最佳表达条件为:诱导时菌液OD600值0.5,加IPTG终浓度0.3mmol/L,诱导时间8h,诱导温度32℃;最佳培养条件为:葡萄糖浓度0,转速230r/min,装液量50mL。

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