Construction of a Chromosome Map for the Phage Group II Staphylococcus aureus Ps55 (1996)
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BibTeX
@MISC{Bannantine96constructionof,
author = {John P. Bannantine and Peter and A. Pattee},
title = {Construction of a Chromosome Map for the Phage Group II Staphylococcus aureus Ps55},
year = {1996}
}
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Abstract
The genome size and a partial physical and genetic map have been defined for the phage group II Staphylococcus aureus Ps55. The genome size was estimated to be 2,771 kb by pulsed-field gel electrophoresis (PFGE) using the restriction enzymes SmaI, CspI, and SgrAI. The Ps55 chromosome map was constructed by transduction of auxotrophic and cryptic transposon insertions, with known genetic and physical locations in S. aureus NCTC 8325, into the Ps55 background. PFGE and DNA hybridization analysis were used to detect the location of the transposon in Ps55. Ps55 restriction fragments were then ordered on the basis of genetic conservation between the two strains. Cloned DNA probes containing the lactose operon (lac) and genes encoding staphylococcal protein A (spa), gamma hemolysin (hlg), and coagulase (coa) were also located on the map by PFGE and hybridization analysis. This methodology enabled a direct comparison of chromosomal organization between NCTC 8325 and Ps55 strains. The chromosome size, gene order, and some of the restriction sites are conserved between the two phage group strains. A physical map of a chromosome consists of an ordered set of restriction fragments that places genomic sequence in the order it exists inside the cell. The resolution of chromosome structure can vary from a macrorestriction map consisting of only a few large, restriction fragments (10, 14, 17, 37) to thousands of small clones of ordered chromosomal DNA (4, 13) to a complete nucleotide sequence (11). The advantages of a physical map over a genetic map are severalfold. Physical maps are faster to construct and require considerably less laborintensive methods. Methods in genetic mapping, such as conjugation, transformation, transduction, and mutagenesis, can be bypassed in physical map construction. Many prokaryotic physical maps have been constructed as a starting point to characterize the chromosome because methods of genetic
Keyphrases
phage group ii staphylococcus physical map chromosome map genetic map genome size restriction fragment cryptic transposon insertion chromosome structure ps55 restriction fragment genetic conservation place genomic sequence restriction site hybridization analysis dna hybridization analysis chromosome size ordered chromosomal dna macrorestriction map consisting ps55 chromosome map direct comparison ps55 background aureus nctc staphylococcal protein lactose operon chromosomal organization laborintensive method ordered set genetic mapping pulsed-field gel electrophoresis complete nucleotide sequence physical location ps55 strain gene order phage group dna probe many prokaryotic physical map physical map construction gamma hemolysin small clone
