Experimental Methods for Studying the BAM Complex in Neisseria meningitidis

Martine P Bos; Ria Tommassen-van Boxtel; J.P.M. Tommassen

doi:10.1007/978-1-4939-2871-2_3

Experimental Methods for Studying the BAM Complex in Neisseria meningitidis

Martine P Bos, Ria Tommassen-van Boxtel, J.P.M. Tommassen

Research output: Contribution to journal › Article › Academic › peer-review

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.

Original language	English
Pages (from-to)	33-49
Number of pages	17
Journal	Methods in Molecular Biology
Volume	1329
DOIs	https://doi.org/10.1007/978-1-4939-2871-2_3
Publication status	Published - 2015

Keywords

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

Access to Document

10.1007/978-1-4939-2871-2_3

Cite this

@article{47d1a1f52d81420485b6353d6bd31171,

title = "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",

author = "Bos, {Martine P} and Boxtel, {Ria Tommassen-van} and J.P.M. Tommassen",

year = "2015",

doi = "10.1007/978-1-4939-2871-2_3",

language = "English",

volume = "1329",

pages = "33--49",

journal = "Methods in Molecular Biology",

issn = "1064-3745",

publisher = "Humana Press",

}

TY - JOUR

T1 - Experimental Methods for Studying the BAM Complex in Neisseria meningitidis

AU - Bos, Martine P

AU - Boxtel, Ria Tommassen-van

AU - Tommassen, J.P.M.

PY - 2015

Y1 - 2015

N2 - 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.

AB - 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.

KW - BAM complex

KW - Expression vector

KW - Gene cloning

KW - Mutagenesis

KW - Neisseria meningitidis

KW - Outer membrane protein

KW - Semi-native SDS-PAGE

U2 - 10.1007/978-1-4939-2871-2_3

DO - 10.1007/978-1-4939-2871-2_3

M3 - Article

C2 - 26427674

SN - 1064-3745

VL - 1329

SP - 33

EP - 49

JO - Methods in Molecular Biology

JF - Methods in Molecular Biology

ER -

Experimental Methods for Studying the BAM Complex in Neisseria meningitidis

Abstract

Keywords

Access to Document

Fingerprint

Cite this