【摘 要】
：
通过Western-blotting筛选出能识别大黄鱼、卵形鲳鲹、青石斑鱼等18种主要经济鱼类血清Ig的大黄鱼(Pseudosciaena crocea)血清Ig单克隆抗体株2H5F4,制备腹水,对纯化后的腹水进行噬菌体肽库淘洗,经过3轮筛选,噬菌体投入产出比为1.65×10-5,噬菌体得到有效富集.挑选7个阳性噬菌体扩增,用M13单链噬菌体DNA提取试剂盒,提取DNA并测序,7个样品中有3个具同
【机 构】
：
福建省农业科学院生物技术研究所, 福州 350003
【出 处】
：
2013海水养殖鱼类病原与病毒控制工程论坛
论文部分内容阅读
通过Western-blotting筛选出能识别大黄鱼、卵形鲳鲹、青石斑鱼等18种主要经济鱼类血清Ig的大黄鱼(Pseudosciaena crocea)血清Ig单克隆抗体株2H5F4,制备腹水,对纯化后的腹水进行噬菌体肽库淘洗,经过3轮筛选,噬菌体投入产出比为1.65×10-5,噬菌体得到有效富集.挑选7个阳性噬菌体扩增,用M13单链噬菌体DNA提取试剂盒,提取DNA并测序,7个样品中有3个具同一序列,其编码的氨基酸序列为：SSTIMER.噬菌体阳性克隆上清与大黄鱼血清Ig的竞争ELISA实验表明,在单抗2H5F4稀释相同倍数情况下,随着噬菌体阳性克隆上清稀释倍数的增加,OD495吸光值呈现逐渐升高的趋势,说明噬菌体所展示的表位可被单抗2H5F4识别,是大黄鱼血清Ig的竞争表位.
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