Sizing of the Leptospira genome by pulsed-field agarose gel electrophoresis

The epidemiological analysis of nosocomial pathogens is a subject of long-standing clinical interest. In recent years, molecular techniques have received increasing attention as a means of analyzing epidemiological interrelationships, thus leading to use of the term molecular epidemiology. Since chromosomal DNA represents a fundamental molecule of cellular identity, there has been particular interest in assessing chromosomal similarity as a measure of epidemiological relatedness.One attractive approach has been to digest chromosomal DNA with restriction enzymes, resulting in a series of different sized fragments that form patterns when comparatively analyzed by agarose gel electrophoresis. In this context, differences in fragment patterns commonly are referred to as restriction-fragment length polymorphisms (RFLPs). Enzymes commonly used to cleave DNA typically recognize numerous sites within the bacterial chromosome. With such enzymes, restriction digestion of DNA from different bacterial isolates results in fragments that are too numerous to compare accurately after conventional agarose gel electrophoresis.

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Physical map of chromosomal and plasmid DNA comprising the genome ofLeptospira interrogans

Zuerner¹

1991

Nucl Acids Res

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The size and physical structure of the Leptospira interrogans genome was characterized using contour-clamped homogenous electric field (CHEF) gel electrophoresis. The L. interrogans genome is approximately 4750 kb in size and is composed of two molecular species of DNA: a 4400 kb chromosome; and a 350 kb plasmid, pLIN1. A physical map of the chromosome was constructed with the restriction enzymes NotI and SfiI. A physical map of pLIN1 was constructed with ApaI, NotI, Sse83871, SgrAI, and SmaI. Both the L. interrogans chromosome and pLIN1 are circular.

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Sizing of the Leptospira genome by pulsed-field agarose gel electrophoresis

Cited by 6 publications

References 13 publications

Differentiation of leptospires of the serogroup Pomona by monoclonal antibodies, pulsed-field gel electrophoresis and arbitrarily primed polymerase chain reaction

Differentiation of leptospires of the serogroup Pomona by monoclonal antibodies, pulsed-field gel electrophoresis and arbitrarily primed polymerase chain reaction

Molecular Epidemiology of Nosocomial Infection: Analysis of Chromosomal Restriction Fragment Patterns by Pulsed-Field Gel Electrophoresis

Physical map of chromosomal and plasmid DNA comprising the genome ofLeptospira interrogans

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