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Use of proteomics and PCR to elucidate changes in protein expression in oral streptococci

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Methods in Cell Science

Abstract

Several techniques are available for the exploration of changes in gene expression and protein repertoire in bacteria due to changes in environment. We have been interested in the changes that S. mutans, a cariogenic oral microorganism, makes in response to growth at low pH values. Two approaches that we have utilized to evaluate alterations in gene expression are the polymerase chain reaction and two-dimensional gel electrophoresis. The use of degenerate primers in PCR amplification of genes by homology-based approaches is included as well as a method for the construction of genetic fusions by splice-overlap extension PCR (SOE-PCR). As genome nucleotide sequences become available for the streptococci, PCR will likely be used more in combination with protein databases, or proteomes, to explore the expression of genes under specific environmental conditions. Therefore, we include a description of two-dimensional gels prepared using protein extracts of cells grown in steady-state under different environmental conditions.

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Authors and Affiliations

  1. Department of Dental Research, University of Rochester, Rochester, New York, USA

    Robert G. Quivey Jr & Roberta C. Faustoferri

  2. Department of Microbiology and Immunology, University of Rochester, Rochester, New York, USA

    Robert G. Quivey Jr & Wendi L. Kuhnert

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  1. Robert G. Quivey Jr
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  2. Wendi L. Kuhnert
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  3. Roberta C. Faustoferri
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Correspondence to Robert G. Quivey Jr.

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Quivey, R.G., Kuhnert, W.L. & Faustoferri, R.C. Use of proteomics and PCR to elucidate changes in protein expression in oral streptococci. Methods Cell Sci 20, 165–179 (1998). https://doi.org/10.1023/A:1009782723759

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