Measuring nitric oxide metabolism in the pathogen Neisseria meningitidis.
This chapter illustrates some of the considerations that need to be made when analyzing nitric oxide (NO) metabolism of the pathogen Neisseria meningitidis. These considerations are pertinent to other bacteria and, in particular, other pathogens. First, because of the small culture volumes that can generally be managed safely, culture experiments are maintained in small volumes. We show a method for simultaneously measuring oxygen and nitric oxide during growth of N. meningitidis in a bioreactor/electrode chamber adapted from commercially available equipment. NO and NO-generating compounds can be used to investigate the impact of NO on N. meningitidis metabolism and gene expression in pure culture. Also, methods are described for analyzing the role of NO during the interaction between N. meningitidis and human macrophage cells that generate NO.
Thomson
Melanie J
MJ
Department of Biology, University of York, Heslington, York.
Stevanin
Tania M
TM
Moir
James W B
JW
D018929:Cell Culture Techniques; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods; D008264:Macrophages / Q000378:metabolism / Q000502:physiology; D008589:Meningococcal Infections / Q000378:metabolism; D008828:Microbiological Techniques / Q000295:instrumentation / Q000379:methods; D008954:Models, Biological; D009345:Neisseria meningitidis / Q000254:growth & development / Q000378:metabolism* / Q000472:pathogenicity; D009569:Nitric Oxide / Q000032:analysis / Q000378:metabolism*; D020030:Nitric Oxide Donors / Q000378:metabolism / Q000494:pharmacology; D009584:Nitrogen / Q000032:analysis / Q000378:metabolism; D016272:Occupational Health; D010100:Oxygen / Q000032:analysis / Q000494:pharmacology; D010802:Phylogeny; D013048:Specimen Handling / Q000379:methods
D020030:Nitric Oxide Donors; D009569:Nitric Oxide; D009584:Nitrogen; D010100:Oxygen