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Abstract:

The classic golden yellow of colonies is caused by carotenoids and develops slowly over several days. is the only species that ferments mannitol anaerobically. The genus contains some 20 distinct species. Two groups of animal-specific, coagulase-variable strains have been granted species status: and . As is generally true of bacteria, staphylococci possess a wide spectrum of variable traits that represent variable genes or variably expressed genes. Study of the genotypic basis of these variable traits has revealed variable genes, such as those carried by plasmids, transposons, and other heterologous genetic elements, and variably expressed genes, such as those belonging to the global regulon. Species-specific surface carbohydrate antigens of and have been identified as teichoic acids. Four different protein hemolysins of are now recognized; all produce clear β-hemolysis, but they differ in RBC species specificity and mechanism of action. A large majority of the toxic shock syndrome (TSST)-1-producing strains have a characteristic biotype that includes sensitivity to typing phage 29 or 52 or both and the presence of chromosomally located resistances to Cd, AsO , and penicillin. During succeeding years, the frequency of resistant strains increased rapidly, and in many clinical settings, multiple antibiotic resistance is now the rule. Even more problematic is the mechanism of pathogenicity of and other opportunistic coagulase-negative species, which have recently equalled and sometimes surpassed as nosocomial pathogens.

Citation: Novick R. 1993. , p 17-33. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch2

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Toxic Shock Syndrome Toxin 1
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Gene Expression and Regulation
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Bacterial Proteins
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Figures

Image of Figure 1
Figure 1

Gram stain of exudate containing intracellular and extracellular staphylococci. Magnification, × 650. (From reference with permission.)

Citation: Novick R. 1993. , p 17-33. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch2
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Figure 2

Scanning electron photomicrograph of in serum-salts broth. (From reference with permission.)

Citation: Novick R. 1993. , p 17-33. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch2
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Figure 3

Dendrogram of phylogenetic relationships based on homologies of 16S rRNA. The scale represents the similarity coefficient (S value; the percentage of shared oligonucleotides in staphylococcal nuclease digests. (From reference with permission.)

Citation: Novick R. 1993. , p 17-33. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch2
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Figure 4

Southern blotting with probes for chromosomal transposons. Lanes 4 through 7 in each panel contain 1 restriction digests of chromosomal DNA from TSS-causing strains that are epidemiologically related. Lanes 1 through 3 contain digests of chromosomal DNA from epidemiologically unrelated TSS strains. (A) Probe specific for the β-lactamase gene; (B) probe specific for Tn (C) probe specific for the TSST-1 gene. (From reference with permission.)

Citation: Novick R. 1993. , p 17-33. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch2
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Figure 5

(A) Negatively stained fragment of rabbit RBC lysed with -toxin. Numerous 10-nm ring-shaped structures are seen over the membrane (arrows). (B) Isolated α-toxin hexamers in detergent solution. (C) Lecithin liposomes carrying reincorporated α-toxin hexamers. The hexamers are seen as stubs along the edge of the liposomal membrane and as rings over the membrane (arrows). Characteristically, liposomes that escape incorporation of the toxin are impermeable to the stain (asterisk). (From reference with permission.)

Citation: Novick R. 1993. , p 17-33. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch2
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Figure 6

Functional genetic map of pT181. Important restriction sites are given with nucleotide positions. Wavy lines represent known transcripts; solid blocks represent known promoters; heavy lines represent reading frames known to encode proteins; lp, putative leader peptide. Functional elements (in counterclockwise order: cop, copy control; ORI, replication origin; RepC, initiator protein coding sequence; Tet, tetracycline resistance determinant (whether there are one or two genes is still uncertain); t, probable termination signal for RS, recombination site A, promoter; Pre, coding sequence; Cmp, competition determinant; RS, recombination site B; Pal A, palindrome A; and countertranscript promoters.

Citation: Novick R. 1993. , p 17-33. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch2
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Figure 7

Map of staphylococcal conjugative plasmid pG0l. Kilobase coordinates begin at 6 o'clock at the single I site (P). Dark arrows are IS-like elements; direction of each arrow indicates the direction of transcription of the single open reading frame on IS, to which these IS-like elements are homologous. Open boxes are antimicrobial resistance genes or the conjugative transfer () region. The arrow at the end of the box designating the trimethoprim resistance gene (Tp) indicates the direction of transcription. The arrow at 5 o'clock indicates the specific RI fragment in which a mobile element encoding penicillinase () inserts. While the element is not present on pG01, its insertion site has been mapped on other homologous conjugative plasmids. Other abbreviations: Gm, gentamicin resistance; Qam, quaternary ammonium-ethidium bromide resistance. Restriction endonuclease cleavage sites are RI (E), II (B), and HI (Bm).

Citation: Novick R. 1993. , p 17-33. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch2
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Figure 8

Maps of cloning vectors useful for (A) pWN1818 ( ). pWN1818 is a promoter-probe vector derived from pA07 ( ), a fusion of () containing the 6.9-kb RI-B fragment of pl258 ( ) to a 0.5-kb segment containing the ColEl origin, ori(E) (12 to 9 o'clock). pC194 (9 to 12 o'clock) was inserted into the unique dIII site of pA07, the promoter was eliminated by 31 digestion, and finally, the pUC18 polylinker (Mc) was inserted as shown ( ). Versions of this vector with stop codons in all three reading frames, with the polylinker in the opposite orientation, or with the Shine-Dalgarno sequence deleted are available. (HindIII), a dIII site that has been filled in and religated; cadA' and asa', truncated and genes; blal, control region with a constitutive mutation. (B) pRN6725 ( ). pRN6725 is a promoter vector derived by inserting the dIII-XI fragment containing the promoter plus two-thirds of the blaZ structural gene into the polylinker region of pRN5543, a pC194 derivative. DNA inserted into the polylinker is transcribed from the promoter, which can be made inducible by providing the repressor in trans. P-bla, -lactamase promoter; blaZ', truncated blaZ; palB, dyad region of unknown function; rep, rep gene of pC194; ori, leading-strand replication origin; cat, Cm gene; palA, lagging-strand replication origin; ori, origin. (C) Protein A fusion vectors pRITl 6 and pRIT21-23. These vectors were constructed by cloning to pEMBL9 a 1.1-kb fragment containing a 3′ truncated gene lacking the coding region for the C-terminal membrane-spanning domain of protein A and containing the pUC18 polylinker sites just 3′ to the fragment. Staphylococcal plasmid pC194 was then inserted between ColEl and regions to give pRIT16, and a transcription termination signal (T) was inserted just past the polylinker region to give pRIT21-23. Cloning to the polylinker in this vector will, if in frame, produce fusion proteins that may be periplasmic in and secreted in (depending on the fused protein). The native promoter is present and is supplemented by the promoter (arrow) ( ). (D) pPL703. This plasmid consists of a 1,250-bp I-II fragment of NCIB8600 DNA inserted between the RI and HI sites of pUB110 by use of ? 21 -bp RI-I fragment from m13mp7 ( ). The promoterless gene resides within the 1,250-bp fragment, and the gene is followed by an efficient transcription termination signal, designated ( ). The pUB 110 portion of pPL703 provides an origin of replication and a neomycin resistance gene (Neo). specifies chloramphenicol acetyltransferase when the gene is transcriptionally activated by inserting a promoter into any of four unique restriction sites 5′ to RI, HI, I, and I. RBS-1, RBS-2, and RBS-3 designate the approximate locations of ribosome-binding sites identified by their complementarity to 26S rRNA. Since the regulatory sequences (→←) are intact, chloramphenicol inducibility is retained. Accordingly, with a promoter-containing derivative of pPL703, cloning of any gene in frame into the coding sequence will result in a chloramphenicol-inducible fusion protein. Reprinted from Lovett et al. ( ) by kind permission from the publishers. Ori, origin.

Citation: Novick R. 1993. , p 17-33. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch2
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Figure 9

Phage 11 map. Restriction sites indicated were mapped by Bachi ( ), Lofdahl et al. ( ), and Novick et al. ( ). The arrow pointing to II-E indicates the ??? site ( ); kbp refers to the circular scale outside the RI map. Early, head, and tail regions were mapped by suppressor-sensitive mutations ( ), and these regions were correlated with the physical map by cloning ( ) (the shaded fragments cause high-frequency transduction when cloned to a plasmid [see text]) and by analysis of the 11::ρ?258 plasmid-phage recombinant 11 ( ). Temperature-sensitive mutations (ts) isolated by this laboratory (N) (unpublished data), by Cohen et al. (sc) ( ), or by Sjostrom and Philipson (sp) ( ) have been localized by cloning. Capital letters represent genes mapped by complementation of mutants ( ); has been mapped by sequencing of the cloned prophage junctions ( ), and has been mapped by cloning ( ).

Citation: Novick R. 1993. , p 17-33. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch2
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Tables

Generic image for table
Table 1

Accessory genetic traits in

Citation: Novick R. 1993. , p 17-33. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch2
Generic image for table
Table 2

Staphylococcal plasmids

Citation: Novick R. 1993. , p 17-33. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch2
Generic image for table
Table 3

Cloning vectors

Citation: Novick R. 1993. , p 17-33. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch2
Generic image for table
Table 4

Transposon delivery vectors

Citation: Novick R. 1993. , p 17-33. In Sonenshein A, Hoch J, Losick R (ed), and Other Gram-Positive Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555818388.ch2

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