Work

Self-made phage libraries with heterologous inserts in the Mtd of Bordetella bronchiseptica

Year: 2012

Type: article

Abstract: Phage display libraries are widely used as tools for identifying, dissecting and optimizing ligands. Development of a simple method to access greater library diversities could expedite and expand the technique. This paper reports progress toward harnessing the naturally occurring diversity generating retroelement used by Bordetella bronchiseptica bacteriophage to alter its tail-fiber protein. Mutagenesis and testing identified four sites amenable to the insertion of <19-residue heterologous peptides within the variable region. Such sites allow auto-generation of peptide libraries surrounded by a scaffold with additional variations. The resultant self-made phage libraries were used successfully for selections targeting anti-FLAG antibody, immobilized metal affinity chromatography microtiter plates and HIV-1 gp41. The reported experiments demonstrate the utility of the major tropism determinant protein of B.bronchiseptica as a natural scaffold for diverse, phage-constructed libraries with heterologous self-made phage libraries.

Source: Protein Engineering Design and Selection

DOI: 10.1093/protein/gzr068

Authors: C. M. Overstreet, T. Z. Yuan, A. M. Levin, C. Kong, J. G. Coroneus, G. A. Weiss

Institution: University of California, Irvine

Language: English

Topic: Bacteriophages and microbial interactions

Subfield: Ecology

Field: Environmental Science

Domain: Physical Sciences

Open Access status: bronze

Funders: National Institutes of Health, National Institute of General Medical Sciences

Award: Engineering Soluble Aggregation-Prone and Membrane-Bound Proteins