PMID: 9642187Jun 27, 1998Paper

Mutagenesis and expression of amo, which codes for ammonia monooxygenase in Nitrosomonas europaea

Journal of Bacteriology
N G HommesD J Arp

Abstract

Nitrosomonas europaea has two copies of the operon encoding ammonia monooxygenase (AMO). The nucleotide sequences of the two copies of amoA were obtained, and they were found to differ by one nucleotide. To determine if both copies of amoA were functional, insertional mutagenesis was performed to inactivate either copy of amoA alone. A DNA cassette containing the lacZ and kan genes inserted into amoA was constructed. Mutagenesis was done by using transformation and homologous recombination to mobilize the cassette into the chromosomal copies of amoA. Mutations were obtained in both copies of amoA. Either copy of amoA was sufficient to support growth when the other copy was disrupted. However, inactivation of one copy of amoA, but not the other, resulted in slower growth. Measurements of ammonia-dependent O2 consumption, which depends on AMO, confirmed that the slower-growing mutant had lower activity while the faster-growing mutant had near wild-type levels of activity. Similarly, as measured by [14C]acetylene label incorporation, there was less active AMO present in the slower-growing mutant than in the faster-growing mutant or in the wild type. Northern blot analysis of transcription likewise showed that the slower-growing mu...Continue Reading

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Citations

Dec 26, 2003·Critical Reviews in Biochemistry and Molecular Biology·Daniel J Arp, Lisa Y Stein
Mar 19, 2005·Extremophiles : Life Under Extreme Conditions·Hisako HirayamaKoki Horikoshi
Jan 31, 2016·Applied and Environmental Microbiology·Kristen BennettMichael R Hyman
Mar 4, 2000·Applied and Environmental Microbiology·B GilbertJ C Murrell
Nov 15, 2000·FEMS Microbiology Reviews·H BotheK Witzel
Oct 24, 1998·FEMS Microbiology Letters·L A Sayavedra-SotoM G Klotz
Oct 31, 2002·Applied and Environmental Microbiology·Linda D RhodesMark S Strom
Jun 9, 2004·Microbiology·Xueming WeiDaniel J Arp
Apr 9, 2008·Applied and Environmental Microbiology·Jeanette M NortonShawn R Starkenburg
Nov 7, 2000·FEMS Microbiology Letters·L Y SteinD J Arp
May 21, 2009·Biological Chemistry·Stefan GilchIngo Schmidt
Mar 9, 2004·Journal of Microbiological Methods·Laia Calvó, L Jesús Garcia-Gil
Oct 14, 2021·Critical Reviews in Biotechnology·Parmita ChawleySheeja Jagadevan

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