Maruyama I N, Maruyama H I, Brenner S
Department of Cell Biology, Scripps Research Institute, La Jolla, CA 92037.
Proc Natl Acad Sci U S A. 1994 Aug 16;91(17):8273-7. doi: 10.1073/pnas.91.17.8273.
This work describes a lambda phage expression system, lambda foo, that produces foreign proteins fused to the surface of the virus particle. The lambda foo vector has multiple cloning sites for the insertion of a foreign DNA fragment and color selection for recombinants. Foreign proteins are fused to the C terminus of a truncated phage tail protein, pV, by a peptide linker. Conditional chain termination allows the assembly and fusion of multisubunit proteins. We have attached the complete Escherichia coli beta-galactosidase and the plant Bauhinia purpurea agglutinin by cloning their genes into the vector. The constructs express functionally active proteins on the phage particle surface and have been purified by affinity chromatography with an antibody for beta-galactosidase and a mucin as a ligand for Bauhinia purpurea agglutinin.
这项工作描述了一种λ噬菌体表达系统——λfoo，它能产生与病毒颗粒表面融合的外源蛋白。λfoo载体具有多个用于插入外源DNA片段的克隆位点以及用于重组体的颜色筛选。外源蛋白通过肽接头与截短的噬菌体尾蛋白pV的C末端融合。条件性链终止允许多亚基蛋白的组装和融合。我们通过将大肠杆菌β-半乳糖苷酶和植物紫羊蹄甲凝集素的基因克隆到该载体中，使其与噬菌体颗粒表面相连。构建体在噬菌体颗粒表面表达具有功能活性的蛋白，并已分别通过用抗β-半乳糖苷酶抗体进行亲和层析以及用粘蛋白作为紫羊蹄甲凝集素的配体进行亲和层析进行了纯化。
