Sternberg N, Hoess R H
Dupont-Merck Pharmaceutical Company, Wilmington, DE 19880-0328.
Proc Natl Acad Sci U S A. 1995 Feb 28;92(5):1609-13. doi: 10.1073/pnas.92.5.1609.
The display of peptides or proteins on the surface of viruses is an important technology for studying peptides or proteins and their interaction with other molecules. Here we describe a display vehicle based on bacteriophage lambda that incorporates a number of features distinct from other currently used display systems. Fusions of peptides or protein domains have been made to the amino terminus of the 11-kDa D protein of the lambda capsid. These fusions assemble onto the viral capsid and appear to be accessible to ligand interactions, based on the ability of a monoclonal antibody to recognize an epitope fused to the D protein on phage heads. To produce large D fusion display libraries and yet avoid the cumbersome task of cloning many fragments into lambda DNA, we have used the Cre-loxP site-specific recombination system in vivo to incorporate plasmids encoding the D fusions into the phage genome. Finally, we show that D fusion proteins can be added in vitro to phage lacking D protein and be assembled onto the viral capsid.
在病毒表面展示肽或蛋白质是研究肽或蛋白质及其与其他分子相互作用的一项重要技术。在此，我们描述一种基于λ噬菌体的展示载体，它具有许多与目前使用的其他展示系统不同的特征。已将肽或蛋白质结构域与λ衣壳11 kDa D蛋白的氨基末端融合。这些融合体组装到病毒衣壳上，基于单克隆抗体识别噬菌体头部与D蛋白融合的表位的能力，似乎可进行配体相互作用。为了构建大型D融合展示文库，同时避免将许多片段克隆到λ DNA中的繁琐任务，我们在体内使用Cre-loxP位点特异性重组系统将编码D融合体的质粒整合到噬菌体基因组中。最后，我们表明D融合蛋白可在体外添加到缺乏D蛋白的噬菌体中，并组装到病毒衣壳上。
