Microbial Biotechnology
Electronic Journal of Biotechnology ISSN: 0717-3458 Vol. 9 No. 5, Issue of October 15, 2006
© 2006 by Pontificia Universidad Católica de Valparaíso -- Chile Received November 3, 2005 / Accepted Abril 4, 2006
DOI: 10.2225/vol9-issue5-fulltext-3  
RESEARCH ARTICLE

Genome analysis of thirteen Colombian clostridial strains by pulsed field gel electrophoresis

Diana Milena Quilaguy Ayure
Bioprocesses and Bioprospecting Group
Instituto de Biotecnología
Universidad Nacional
Ciudad Universitaria, Edificio Manuel Ancizar
Bogotá, Colombia
Tel: 571 3165000. Ext.16970/71
Fax: 571 3165415
E-mail: dmquilaguya@unal.edu.co

Zulma Rocío Suárez Moreno*
Bioprocesses and Bioprospecting Group
Instituto de Biotecnología
Universidad Nacional
Ciudad Universitaria, Edificio Manuel Ancizar
Bogotá, Colombia
Tel: 571 3165000. Ext.16972/70
Fax: 571 3165415
E-mail: zrsuarezm@unal.edu.co

Fabio Ancizar Aristizábal Gutierrez
Bioprocesses and Bioprospecting Group
Instituto de Biotecnología
Universidad Nacional
Ciudad Universitaria
Bogotá, Colombia
Tel: 571 3165000. Ext.14643
Fax: 571 3165415
E-mail: faaristizabalg@unal.edu.co

Jose Mauricio Bernal Morales
Bioprocesses and Bioprospecting Group
Instituto de Biotecnología
Universidad Nacional
Ciudad Universitaria, Edificio Manuel Ancizar
Bogotá, Colombia
Tel: 571 3165000. Ext.16970/71
Fax: 571 3165415
E-mail: jmbernalmo@unal.edu.co

 Dolly Montoya Castaño
Bioprocesses and Bioprospecting Group
Instituto de Biotecnología
Universidad Nacional
Ciudad Universitaria, Edificio Manuel Ancizar
Bogotá, Colombia
Tel: 571 3165000. Ext.16952
Fax: 571 3165415
E-mail: dmontoyac@unal.edu.co

*Corresponding author

Financial support: Colciencias, Universidad Nacional de Colombia.

Keywords: Clostridium, genome size, PFGE, pSOL1.

Abbreviations:

AFLP: amplified fragment length polymorphism
ATCC: American Type Culture Collection
bp: base pairs
(CHEF)-PFGE: contour-clamped homogeneous electric field
EC: lysis buffer
EDTA: ethylenediaminetetracetic acid
ES buffer: EDTA–sarcosine buffer
ESP: EDTA-sarcosine-proteinase buffer
ET buffer: EDTA-Tris Buffer (Tris-HCl 10 mM, EDTA 100 mM)
IBUN: Instituto de Biotecnología de la Universidad Nacional
Mbp: mega base pairs
OD: optical density
ORF: open reading frame
PCR: polymerase chain reaction
PFGE: pulsed field gel electrophoresis
PMSF: phenylmethylsulphonyl fluoride
RCM: reinforced clostridial medium
RPM: Revolutions Per Minute
TBE: Tris-borate-EDTA buffer
TE: Tris-EDTA buffer (Tris-HCl 10 mM, EDTA 1 mM)

Abstract
Full Text

Pulsed field gel electrophoresis was used for estimating the size of the genome and evaluating the presence of megaplasmids in 13 native Colombian solventogenic Clostridium strains. DNA preparation and purification were optimised for obtaining differentiated restriction fragments in electrophoresis. Genomic DNA was digested with ApaI, Eco52I, SmaI and XhoI enzymes. Estimated genome size for native strains ranged from 4.0 to 4.2 mega base pairs. Larger sized plasmids were detected and the presence of genes related to megaplasmid pSOL1 was determined by polymerase chain reaction. adc gene region amplification suggested that genes related to solventogenesis in native strains may be located in an extra-chromosomal element. Determining genome size provides useful information aimed at enhancing native strains' solvent production.
Supported by UNESCO / MIRCEN network
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