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EXPRESSION CLONING OF CDNA BY PHAGE DISPLAY SELECTION

机译:通过噬菌体显示选择对CDNA进行表达克隆

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Expression cloning of a mouse kappa chain fragment has been achieved from a cDNA library by display of expressed proteins on filamentous phage and affinity selection for binding to anti-mouse Fab antibodies. Expressed proteins were anchored to the phage coat by a synthetic, anti-parallel leucine zipper, which had been selected from a semi-randomized zipper library for the ability to connect a test protein to phage. From a library of 4 x 10(6) transformants, two separate clones displaying different size cDNA inserts were recovered after four selection rounds. These results further demonstrate the utility of phage display for cDNA expression cloning.
机译:通过在丝状噬菌体上展示表达的蛋白质并选择与抗小鼠Fab抗体结合的亲和力,已经从cDNA文库中完成了小鼠κ链片段的表达克隆。表达的蛋白质通过合成的反平行亮氨酸拉链固定在噬菌体外壳上,该拉链选自半随机的拉链文库,具有将测试蛋白质连接到噬菌体的能力。从四个x 10(6)转化子的库中,经过四轮选择后,获得了显示大小不同的cDNA插入片段的两个单独的克隆。这些结果进一步证明了噬菌体展示对cDNA表达克隆的实用性。

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