DISPLAY OF PEPTIDES AND PROTEINS ON THE SURFACE OF BACTERIOPHAGE-LAMBDA

被引:149
作者
STERNBERG, N
HOESS, RH
机构
[1] Dupont-Merck Pharmaceutical Company, Experimental Station E328/B33, Wilmington
关键词
PHAGE DISPLAY; BIOPLANNING; D PROTEIN; SITE-SPECIFIC RECOMBINATION; LIBRARIES;
D O I
10.1073/pnas.92.5.1609
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
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.
引用
收藏
页码:1609 / 1613
页数:5
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