Mikawa Y G, Maruyama I N, Brenner S
Department of Cell Biology, Scripps Research Institute, La Jolla CA 92037, USA.
J Mol Biol. 1996 Sep 13;262(1):21-30. doi: 10.1006/jmbi.1996.0495.
We have developed plasmid and phage vectors for the display of foreign proteins on the surface of bacteriophage lambda capsid by modifying the D gene which encodes the major head protein gpD. The vectors have multiple cloning sites, and permit colour selection and conditional chain termination for recombinants. Displayed proteins can be fused to either the N or C terminus of gpD by a peptide linker. The conditional chain termination scheme, via a host Escherichia coli suppressor activity, allows the fusion and assembly of homomultimeric proteins as well as control of the number of fusion proteins per phage particle. We have successfully displayed beta-lactamase, IgG-binding domains of the Staphylococcus aureus protein A, and beta-galactosidase by cloning the genes into the vector. The constructs express functionally active proteins fused to gpD that assemble into phage particles. These results suggest that gpD may be fused to many other peptides and proteins at their N or C terminus and the fusion products may be accessible on the surface of bacteriophage lambda particles.
我们通过修饰编码主要头部蛋白gpD的D基因，开发了用于在噬菌体λ衣壳表面展示外源蛋白的质粒和噬菌体载体。这些载体具有多个克隆位点，并允许对重组体进行颜色筛选和条件性链终止。展示的蛋白可通过肽接头与gpD的N端或C端融合。通过宿主大肠杆菌抑制活性的条件性链终止方案，允许同源多聚体蛋白的融合和组装，以及控制每个噬菌体颗粒上融合蛋白的数量。我们通过将基因克隆到载体中，成功展示了β-内酰胺酶、金黄色葡萄球菌蛋白A的IgG结合结构域和β-半乳糖苷酶。构建体表达与gpD融合的功能活性蛋白，这些蛋白组装成噬菌体颗粒。这些结果表明，gpD可在其N端或C端与许多其他肽和蛋白融合，并且融合产物可能在噬菌体λ颗粒表面可及。
