Pulsed-Field Separations: Continued Evolution

Eby, Michael J.

doi:10.1038/nbt0390-243

“…The influence of these parameters has been investigated systematically by Cantor et al (1988), Mathew et al (1988a,b,c) and Doggett et al (1992). During the past years, several different PFGE techniques have been published, like Field-Inversion Gel Electrophoresis (FIGE; Carle et al 1986); ContourClamped Homogeneous Electric Field (CHEF; Chu et al 1986); Transverse Alternating Field Electrophoresis (TAFE; Gardiner and Patterson 1988); Rotating System (RGE or RFE; for a review see Eby 1990); Secondary Pulsed Field Gel Electrophoresis (Zhang et al 1991) and Zero-Integrated-Field Electrophoresis (ZIFE; Noolandi and Tunnel 1992). All these techniques have proven useful for electrophoretic karyotyping of yeasts.…”

Section: Introduction and Theorymentioning

confidence: 99%

Electrophoretic Karyotyping of Yeasts

Zimmermann

¹

,

Fournier²

1996

Nonconventional Yeasts in Biotechnology

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“…(34). Unable to travel in a straight path, due to the size of the DNA, the DNA would elongate and compact allowing it to move through the gaps in the gel matrix (61,214); this however caused the DNA to travel in snake like paths through the gel in an effort to find pores of the correct size (214). PFGE overcame this by pulsing the electrical current from different points around the gel at given time intervals, allowing the previously cut DNA sample to migrate in a straight line (34,214).…”

Section: Flaa Pcr Rflpmentioning

confidence: 99%

“…The most common PFGE method used is contour-clamped homogeneous electric field electrophoresis (CHEF) (34,61). CHEF was developed by Chu et al (1986) to resolve large DNA molecules (34).…”

Section: Flaa Pcr Rflpmentioning

confidence: 99%

“…CHEF was developed by Chu et al (1986) to resolve large DNA molecules (34). The CHEF-PFGE current field is arranged in a hexagonal shape, with electrodes delivering pulses from an angle of 120 ( 61).The current pulsed at 120˚ angles pushes larger DNA molecules through the gel matrix in a uniform manner, allowing better resolution (61) than other electric fields generated using with 60-90˚ angles (34).…”

Section: Flaa Pcr Rflpmentioning

confidence: 99%

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Comparative Analysis of New Zealand Campylobacter Isolates Using MLST, PFGE and flaA PCR RFLP Genotyping

McTavish¹

0

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<p>Campylobacter jejuni and Campylobacter coli are the most commonly identified sources of campylobacteriosis in New Zealand, yet little is known about the distribution of genotypes within the respective population structures. Using multi-locus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE) and flaA genotyping, the current study identified the distribution of genotypes within New Zealand C. jejuni and C. coli isolates from an outbreak situation, as well as isolates present in the ESR Campylobacter collection. Although the most commonly identified MLST genotypes were similar to international genotypes, a number of internationally rare, or unique to New Zealand genotypes were observed. One rare dominant genotype, ST-474, arising from a point source outbreak, was found to cause a large proportion of human campylobacteriosis cases in New Zealand. A unique cluster of New Zealand genotypes were isolated only from river water, identifying a potentially water adapted C. jejuni strain. Frequent homologous recombination and horizontal gene transfer events were identified within the seven housekeeping genes characterised in the New Zealand sample and the MLST C. jejuni/C. coli database. The identified genetic instability within the current study questions the legitimacy of bacterial species boundaries, especially when examining closely related species such as C. jejuni and C. coli.</p>

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“…(34). Unable to travel in a straight path, due to the size of the DNA, the DNA would elongate and compact allowing it to move through the gaps in the gel matrix (61,214); this however caused the DNA to travel in snake like paths through the gel in an effort to find pores of the correct size (214). PFGE overcame this by pulsing the electrical current from different points around the gel at given time intervals, allowing the previously cut DNA sample to migrate in a straight line (34,214).…”

Section: Flaa Pcr Rflpmentioning

confidence: 99%

Comparative Analysis of New Zealand Campylobacter Isolates Using MLST, PFGE and flaA PCR RFLP Genotyping

McTavish¹

0

View full text Add to dashboard Cite

<p>Campylobacter jejuni and Campylobacter coli are the most commonly identified sources of campylobacteriosis in New Zealand, yet little is known about the distribution of genotypes within the respective population structures. Using multi-locus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE) and flaA genotyping, the current study identified the distribution of genotypes within New Zealand C. jejuni and C. coli isolates from an outbreak situation, as well as isolates present in the ESR Campylobacter collection. Although the most commonly identified MLST genotypes were similar to international genotypes, a number of internationally rare, or unique to New Zealand genotypes were observed. One rare dominant genotype, ST-474, arising from a point source outbreak, was found to cause a large proportion of human campylobacteriosis cases in New Zealand. A unique cluster of New Zealand genotypes were isolated only from river water, identifying a potentially water adapted C. jejuni strain. Frequent homologous recombination and horizontal gene transfer events were identified within the seven housekeeping genes characterised in the New Zealand sample and the MLST C. jejuni/C. coli database. The identified genetic instability within the current study questions the legitimacy of bacterial species boundaries, especially when examining closely related species such as C. jejuni and C. coli.</p>

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Pulsed-Field Separations: Continued Evolution

Cited by 7 publications

References 14 publications

Electrophoretic Karyotyping of Yeasts

Electrophoretic Karyotyping of Yeasts

Comparative Analysis of New Zealand Campylobacter Isolates Using MLST, PFGE and flaA PCR RFLP Genotyping

Comparative Analysis of New Zealand Campylobacter Isolates Using MLST, PFGE and flaA PCR RFLP Genotyping

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