Automated Laboratory Growth Assessment and Maintenance of Azotobacter vinelandii

Brooke M. Carruthers, Amanda K. Garcia, Alex Rivier, Betul Kacar

Research output: Contribution to journalArticlepeer-review

Abstract

Azotobacter vinelandii (A. vinelandii) is a commonly used model organism for the study of aerobic respiration, the bacterial production of several industrially relevant compounds, and, perhaps most significantly, the genetics and biochemistry of biological nitrogen fixation. Laboratory growth assessments of A. vinelandii are useful for evaluating the impact of environmental and genetic modifications on physiological properties, including diazotrophy. However, researchers typically rely on manual growth methods that are oftentimes laborious and inefficient. We present a protocol for the automated growth assessment of A. vinelandii on a microplate reader, particularly well-suited for studies of diazotrophic growth. We discuss common pitfalls and strategies for protocol optimization, and demonstrate the protocol's application toward growth evaluation of strains carrying modifications to nitrogen-fixation genes.

Original languageEnglish (US)
Article numbere57
JournalCurrent Protocols
Volume1
Issue number3
DOIs
StatePublished - Mar 2021

Keywords

  • Azotobacter vinelandii
  • bacterial growth
  • microplate reader
  • nitrogen fixation
  • nitrogenase

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Pharmacology, Toxicology and Pharmaceutics(all)
  • Neuroscience(all)
  • Immunology and Microbiology(all)
  • Health Informatics
  • Medical Laboratory Technology
  • Medicine(all)

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