Experimental Methods for Studying the BAM Complex in Neisseria meningitidis

Abstract

Neisseria meningitidis is a human pathogen. It is intensively studied for host-pathogen interactions and vaccine development. However, its favorable growth properties, genetic accessibility, and small genome size also make it an excellent model organism for studying fundamental biological processes, such as outer membrane biogenesis. Indeed, the first component of the assembly machinery for outer-membrane proteins, the BAM complex, was identified in N. meningitidis. Here, we describe protocols to inactivate chromosomal genes and to express genes from a well-controlled promoter on a plasmid in N. meningitidis. Together, these protocols can be used, for example, to deplete cells from essential components of the BAM complex. We also describe a simple, gel-based assay to assess the proper functioning of the BAM complex in vivo.

Keywords: BAM complex; Expression vector; Gene cloning; Mutagenesis; Neisseria meningitidis; Outer membrane protein; Semi-native SDS-PAGE.