Genomic analysis of Clostridium botulinum group II by pulsed -field gel electrophoresis

Pulsed-field gel electrophoresis (PFGE) was optimized for genomic analyses of Clostridium botulinum (non-proteolytic) group II. DNA degradation problems caused by extracellular DNases were overcome by fixation of cells with formaldehyde prior to isolation. A rapid (4-h) in situ DNA isolation method...

Full description

Saved in:
Bibliographic Details
Published in:Applied and Environmental Microbiology 1998-02, Vol.64 (2), p.703-708
Main Authors: Hielm, S, Bjorkroth, J, Hyytia, E, Korkeala, H
Format: Article
Language:English
Subjects:
Bacteria
Bacterial Typing Techniques
Bacteriological methods and techniques used in bacteriology
Bacteriology
Biological and medical sciences
Botulism
CLOSTRIDIUM BOTULINUM
Clostridium botulinum - classification
Clostridium botulinum - genetics
Deoxyribonucleases - metabolism
DNA, Bacterial - analysis
Electrophoresis, Gel, Pulsed-Field
Epidemiology
Food Microbiology
Fundamental and applied biological sciences. Psychology
Genetics
Genome, Bacterial
Microbiology
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Pulsed-field gel electrophoresis (PFGE) was optimized for genomic analyses of Clostridium botulinum (non-proteolytic) group II. DNA degradation problems caused by extracellular DNases were overcome by fixation of cells with formaldehyde prior to isolation. A rapid (4-h) in situ DNA isolation method was also assessed and gave indistinguishable results. Genomic DNA from 21 strains of various geographical and temporal origins was digested with 15 rare-cutting restriction enzymes. Of these, ApaI, MluI, NruI, SmaI, and XhoI gave the most revealing PFGE patterns, enabling strain differentiation. Twenty strains yielded PFGE patterns containing 13 pulsotypes. From summation of MluI, SmaI, and XhoI restriction fragments, the genome size of C. botulinum group II was estimated to be 3.6 to 4.1 Mb (mean +/- standard deviation = 3,890 +/- 170 kb). The results substantiate that after problems due to DNases are overcome, PFGE analysis will be a reproducible and highly discriminating epidemiological method for studying C. botulinum group II at the molecular level
ISSN:0099-2240
1098-5336
DOI:10.1128/aem.64.2.703-708.1998