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Chapter 34 : The Genetic Map of Staphylococcus aureus

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Abstract:

Only when Lindberg and coworkers reported transformation inNCTC 8325, and when some functionally unrelated genes were shown to be cotransformable, was it feasible to attempt to develop a genetic map of the chromosome. Transformation with high-molecular-weight DNA confirmed the orientation and linkage of two of the major linkage groups previously characterized by transformation. However, continuity of these linkage groups on a single circular genetic map, inferred by protoplast fusion, remained unsubstantiated. Furthermore, the genetic map was based almost solely on NCTC 8325, a phage group III strain of . The availability of methods for the construction of physical maps of prokaryotic chromosomes based on pulsed-field agarose gel electrophoresis (PFGE) and DNA hybridization, combined with existing libraries of genetically mapped insertions of transposons , , , and , allowed the construction of a physical map of NCTC 8325 that was correlated with the genetic map. At present, the most efficient way t o identify the chromosomal locus occupied by a new marker of interest is by a combination of physical mapping by PFGE with -, -, and -digested chromosomal DNA and DNA hybridization with an appropriate probe.

Citation: Pattee P. 1993. The Genetic Map of Staphylococcus aureus, p 489-496. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch34

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Agarose Gel Electrophoresis
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Pulsed-Field Gel Electrophoresis
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Chromosomal DNA
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Figure 1

Chromosome map of The identity of individual markers is in Table 1 . The order of the majority of the markers shown is based on genetic analysis and DNA hybridization analysis of 1, 1, and Al restriction fragments resolved by PFGE. Among these markers, Ω11, Ω402, Ω420, Ω500, Ω1005, Ω401, Ω2004, and have not been physically mapped, and their map locations are based only on multifactorial transformation analyses. The silent insertions of Tn shown to be near and near (on the Al F fragment) are readily cotransducible with and respectively. The order of markers shown within parentheses is not known. The entire chromosome is about 2,800 kb, and the individual fragments are drawn approximately to scale. The gaps in the Al map reflect regions that may be occupied by smaller fragments not located on the Al physical map (Al-G, -J, and -K, representing a total of about 300 kb). There are at least two 1 fragments representing about 100 kb, and three 1 fragments representing about 50 kb, that also have not been located on the map.

Citation: Pattee P. 1993. The Genetic Map of Staphylococcus aureus, p 489-496. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch34
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Tables

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Table 1

Genetic markers of the chromosome of S.

Citation: Pattee P. 1993. The Genetic Map of Staphylococcus aureus, p 489-496. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch34

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