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

@article{citeulike:3912293, abstract = {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.}, author = {Hielm, S. and Bj\"{o}rkroth, J. and Hyyti\"{a}, E. and Korkeala, H.}, citeulike-article-id = {3912293}, citeulike-linkout-0 = {http://view.ncbi.nlm.nih.gov/pubmed/9464411}, citeulike-linkout-1 = {http://www.hubmed.org/display.cgi?uids=9464411}, issn = {0099-2240}, journal = {Applied and environmental microbiology}, keywords = {botulinum, phylogeny, typing}, month = {February}, number = {2}, pages = {703--708}, posted-at = {2009-01-20 15:06:10}, priority = {2}, title = {Genomic analysis of Clostridium botulinum group II by pulsed-field gel electrophoresis.}, url = {http://view.ncbi.nlm.nih.gov/pubmed/9464411}, volume = {64}, year = {1998} }

TY - JOUR ID - citeulike:3912293 L3 - citeulike-article-id:3912293 N2 - 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. IS - 2 JF - Applied and environmental microbiology SN - 0099-2240 EP - 708 TI - Genomic analysis of Clostridium botulinum group II by pulsed-field gel electrophoresis. VL - 64 SP - 703 KW - botulinum KW - phylogeny KW - typing AU - Hielm, S AU - Björkroth, J AU - Hyytiä, E AU - Korkeala, H PY - 1998/02// UR - http://view.ncbi.nlm.nih.gov/pubmed/9464411 ER -