Book chapter
Characterizing Bacterial Gene Expression in Nitrogen Cycle Metabolism with RT-qPCR
Methods in Enzymology, pp.345-372
Elsevier Science & Technology
2011
Handle:
https://hdl.handle.net/2376/107057
PMID: 21514471
Abstract
Recent advances in DNA sequencing have greatly accelerated our ability to obtain the raw information needed to recognize both known and potential novel modular microbial genomic capacity for nitrogen metabolism. With PCR-based approaches to quantifying microbial mRNA expression now mainstream in most laboratories, researchers can now more efficiently propose and test hypotheses on the contributions of individual microbes to the biological accessibility of nitrogen upon which all other life depends. We review known microbial roles in these key nitrogen transformations, and describe the necessary steps in carrying out relevant gene expression studies. An example experimental design is then provided characterizing
Nitrosococcus oceani mRNA expression in cultures responding to ammonia. The approach described, that of assessing microbial genome inventory and testing putative modular gene expression by mRNA quantification, is likely to remain an important tool in understanding individual microbial contributions within microbial community activities that maintain the Earth's nitrogen balance.
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Details
- Title
- Characterizing Bacterial Gene Expression in Nitrogen Cycle Metabolism with RT-qPCR
- Creators
- James E Graham - Department of Biology, University of Louisville, Louisville, Kentucky, USANicholas B Wantland - Department of Microbiology and Immunology, University of Louisville, Louisville, Kentucky, USAMark Campbell - Department of Biology, University of Louisville, Louisville, Kentucky, USAMartin G Klotz - Department of Biology, University of Louisville, Louisville, Kentucky, USA
- Publication Details
- Methods in Enzymology, pp.345-372
- Academic Unit
- Molecular Biosciences, School of
- Publisher
- Elsevier Science & Technology
- Identifiers
- 99900547095201842
- Language
- English
- Resource Type
- Book chapter