This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Lanoil, B. D. Articles by Giovannoni, S. J. Search for Related Content PubMed PubMed Citation Articles by Lanoil, B. D. Articles by Giovannoni, S. J. Agricola Articles by Lanoil, B. D. Articles by Giovannoni, S. J.

 Previous Article  |  Next Article 

Appl. Environ. Microbiol., Mar 1997, 1118-1123, Vol 63, No. 3
Copyright © 1997, American Society for Microbiology

Identification of bacterial cells by chromosomal painting

BD Lanoil and SJ Giovannoni
Molecular and Cellular Biology Program, Oregon State University, Corvallis 97331, USA.

Chromosomal painting is a technique for the microscopic localization of genetic material. It has been applied at the subcellular level to identify regions of eukaryotic chromosomes. Here we describe the development of bacterial chromosomal painting (BCP), a related technology for the identification of bacterial cells. Purified genomic DNAs from six bacterial strains were labeled by nick translation with the fluorochrome Fluor-X, Cy3, or Cy5. The average size of the labeled fragments was ca. 50 to 200 bp. The probes were hybridized to formaldehyde-fixed microbial cells attached to slides and visualized by fluorescence microscopy. In reciprocal comparisons, distantly related members of the class Proteobacteria (Escherichia coli and Oceanospirillum linum), different species of the genus Bacillus (B. subtilis and B. megaterium), and different serotypes of the subspecies Salmonella choleraesuis subsp. choleraesuis (serotype typhimurium LT2 and serotype typhi Ty2) could easily be distinguished. A combination of two probes, each labeled with a different fluorochrome, was used successfully to simultaneously identify two cell types in a mixture. Lysozyme treatment was required for the identification of Bacillus spp., and RNase digestion and pepsin digestion were found to enhance signal strength and specificity for all cell types tested. Chromosome in situ suppression, a technique that removes cross-hybridizing fragments from the probe, was necessary for the differentiation of the Salmonella serotypes but was not required to distinguish the more distantly related taxa. BCP may have applications in diverse branches of microbiology where the objective is the identification of bacterial cells.

This article has been cited by other articles:

• Maruyama, F., Kenzaka, T., Yamaguchi, N., Tani, K., Nasu, M. (2005). Visualization and Enumeration of Bacteria Carrying a Specific Gene Sequence by In Situ Rolling Circle Amplification. Appl. Environ. Microbiol. 71: 7933-7940 [Abstract] [Full Text]  
• Kenzaka, T., Tamaki, S., Yamaguchi, N., Tani, K., Nasu, M. (2005). Recognition of Individual Genes in Diverse Microorganisms by Cycling Primed In Situ Amplification. Appl. Environ. Microbiol. 71: 7236-7244 [Abstract] [Full Text]  
• Jaspers, E., Overmann, J. (2004). Ecological Significance of Microdiversity: Identical 16S rRNA Gene Sequences Can Be Found in Bacteria with Highly Divergent Genomes and Ecophysiologies. Appl. Environ. Microbiol. 70: 4831-4839 [Abstract] [Full Text]  
• Maruyama, F., Kenzaka, T., Yamaguchi, N., Tani, K., Nasu, M. (2003). Detection of Bacteria Carrying the stx2 Gene by In Situ Loop-Mediated Isothermal Amplification. Appl. Environ. Microbiol. 69: 5023-5028 [Abstract] [Full Text]  
• Pernthaler, A., Preston, C. M., Pernthaler, J., DeLong, E. F., Amann, R. (2002). Comparison of Fluorescently Labeled Oligonucleotide and Polynucleotide Probes for the Detection of Pelagic Marine Bacteria and Archaea. Appl. Environ. Microbiol. 68: 661-667 [Abstract] [Full Text]  
• Kisand, V., Cuadros, R., Wikner, J. (2002). Phylogeny of Culturable Estuarine Bacteria Catabolizing Riverine Organic Matter in the Northern Baltic Sea. Appl. Environ. Microbiol. 68: 379-388 [Abstract] [Full Text]  
• Urbach, E., Vergin, K. L., Giovannoni, S. J. (1999). Immunochemical Detection and Isolation of DNA from Metabolically Active Bacteria. Appl. Environ. Microbiol. 65: 1207-1213 [Abstract] [Full Text]  
• Nübel, U., Garcia-Pichel, F., Kühl, M., Muyzer, G. (1999). Quantifying Microbial Diversity: Morphotypes, 16S rRNA Genes, and Carotenoids of Oxygenic Phototrophs in Microbial Mats. Appl. Environ. Microbiol. 65: 422-430 [Abstract] [Full Text]  
• Ogunseitan, O. A. (1998). Protein Method for Investigating Mercuric Reductase Gene Expression in Aquatic Environments. Appl. Environ. Microbiol. 64: 695-702 [Abstract] [Full Text]