Work

A New Suite of Allelic-Exchange Vectors for the Scarless Modification of Proteobacterial Genomes

Year: 2019

Type: article

Abstract: Targeted modification of bacterial genomes is critical for genetic analysis of microorganisms. Allelic exchange is a technique that relies on homologous recombination to replace native loci with engineered sequences. However, current allelic-exchange vectors often enable only weak selection for successful homologous recombination. We developed a suite of new allelic-exchange vectors, pTOX, which were validated in several medically important proteobacteria. They encode visible nonfluorescent chromoproteins that enable easy identification of colonies bearing integrated vectors and permit stringent selection for the second step of homologous recombination. We demonstrate the utility of these vectors by using them to investigate the effect of inactivation of Serratia marcescens peptidoglycan amidohydrolases on β-lactam antibiotic resistance.

Source: Applied and Environmental Microbiology

DOI: 10.1128/aem.00990-19

Authors: Jacob E. Lazarus, Alyson R. Warr, Carole J. Kuehl, Rachel T. Giorgio, Brigid M. Davis, Matthew K. Waldor

Institutions: Harvard University, Massachusetts General Hospital, Brigham and Women's Hospital, Howard Hughes Medical Institute

Language: English

Topic: Bacterial Genetics and Biotechnology

Subfield: Genetics

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: green

Funders: Howard Hughes Medical Institute, Harvard Catalyst, National Institutes of Health, National Institute of Allergy and Infectious Diseases