Roberts B L, Markland W, Ladner R C
Genzyme Corporation, Framingham, Massachusetts 01701, USA.
Methods Enzymol. 1996;267:68-82. doi: 10.1016/s0076-6879(96)67006-2.
This chapter described the preparation and fractionation of libraries of M13 phage displaying proteins as fusions to the major coat protein. High titer (10(13) pfu/ml) phage libraries can readily be generated using a single vector and the level of display surpasses that of gene III fusion phage. Since the synthetic VIII fusion gene can be customized, this system should provide the flexibility required to construct phage libraries displaying a variety of different peptides and proteins and to select variants possessing the highest affinity for target molecules of a diverse chemical nature.
本章描述了M13噬菌体文库的制备和分级分离,该文库展示了与主要衣壳蛋白融合的蛋白质。使用单一载体可以轻松生成高滴度(10¹³ pfu/ml)的噬菌体文库,其展示水平超过了基因III融合噬菌体。由于合成的VIII融合基因可以定制,该系统应提供构建展示各种不同肽和蛋白质的噬菌体文库以及选择对具有不同化学性质的靶分子具有最高亲和力的变体所需的灵活性。