Molecular Genetic Studies of Relationships Among Mycoplasma, L‐forms and Bacteria*

McGee, Zell A.; Rogul, Marvin; Wittler, Ruth G.

doi:10.1111/j.1749-6632.1967.tb27639.x

Cited by 55 publications

(39 citation statements)

References 24 publications

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“…Capri, M. bovigenitalium, M . arginini and Acholeplasma laidlawii) our results are in good agreement with earlier investigations (Neimark & Peni, 1965;McGee, Rogul & Wittler, 1967;Morowitz, Bode & Kirk, 1967;Neimark, 1967;Chelton, Jones & Walker, 1968;Kelton & Mandel, 1969;Williams, Wittler & Burris, 1969;Gourlay & Leach, I 970; Neimark, I 970). Discrepancies were, however, found as regards Mycoplasma bovirliinis and both subspecies of M .…”

Section: Discussionsupporting

confidence: 82%

Bovine Mycoplasmas: Genome Size and Base Composition of DNA

Askaa

Christiansen

Ernø

1973

Journal of General Microbiology

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Within the order Mycoplasmatales guanine plus cytosine contents and genome sizes have been determined for 17 strains representing 15 bovine and two ovine species, subspecies or serogroups. The guanine plus cytosine contents were found to vary between 24-4 and 32.9 %. The genome sizes were 4.0 to 5-6 x 1oS daltons for 14 sterol-requiring strains and 0.99 to 1.1 x 109 daltons for three non-sterol-requiring strains. These results support earlier findings that members of the genus Acholeplasma (family Acholeplasmataceae) have genome sizes about twice those of the genus Mycoplasma (family Mycoplasmataceae). The purpose of this work was to determine the GC contents and the genome sizes of type or reference strains of all known mycoplasma species, subspecies and serogroups of bovine source. The type strains (PG2 and PG3) of two ovine organisms (Mycoplasma agalactiae subsp. agalactiae and M. mycoides subsp. Capri) were included in the study because of serological relatedness to the bovine strains ( M . agalactiae subsp. bovis (Donetta) and M . mycoides subsp. mycoides (PGI), respectively). METHODSThe mycoplasmas investigated (Table I) The substrates used for cultivation were in most cases Bacto Heart Infusion Broth (Difco), supplemented with 15 % horse serum or I % PPLO Serum Fraction, 8 % fresh yeast extract, 0.003 % phenol red, 0.08 % thallium acetate and penicillin (400 i.u./ml). Some strains were also supplied with 0.8 % glucose (PG49, PG50, B107PA7 PGI, PG3) or I % arginine (B139P7 B I~~P , ~230). Mycoplasma dispar was cultivated in FF I1 medium (Friis, 1972), with minor modifications; it comprised Bacto Brain Heart Infusion Broth (Difco, Detroit, Michigan, U.S.A.),

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Section: Discussionsupporting

confidence: 82%

Bovine Mycoplasmas: Genome Size and Base Composition of DNA

Askaa

Christiansen

Ernø

1973

Journal of General Microbiology

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“…Thus, M. hominis, the only species that does not ferment carbohydrates and whose antigens have been studied, has antigenic determinants which are different from those of the carbohydrate-fermenting species. What is known of Mycoplasma species antigens suggests that the mycoplasmas comprise a heterogeneous group, a supposition compatible with the pronounced differences in the DNA base ratios within this genus (Neimark, 1967;McGee, Rogul & Wittler, 1967).…”

Section: Discussionmentioning

confidence: 88%

The antigens ofMycoplasma hominis

Hollingdale

Lemcke

1969

Epidemiol. Infect.

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SUMMARYA genital strain ofMycoplasma hominiswas fractionated by differential centrifugation after disruption of the cells by alternate cycles of freezing and thawing, by gas cavitation under nitrogen, or by ultrasonic treatment.Antigens of the cell membrane were distinct from those in the soluble cell contents, judging from metabolic inhibition (MI), indirect haemagglutination (IHA), gel-diffusion and immunoelectrophoresis tests, and by the antibody response in rabbits inoculated with these fractions. The antigens which gave rise to MI and IHA antibody were located in the cell membrane.Extraction of whole cells ofM. hominisby various chemical methods suggested that the active components were protein in nature and that there was no lipid hapten, as inM. pneumoniae, or polysaccharide, as inM. mycoidesvar.mycoides.This work was partly supported by a Medical Research Council grant, which provided the remuneration of MRH.We thank Professor R. A. Kekwick and Dr J. M. Creeth of the Biophysics Department of this Institute for their advice on the processing of plasma and density gradient centrifugation, and Mr F. C. Belton of the Microbiological Research Establishment, Porton, Wilts, for growing a 50 litre batch ofM. hominis.

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“…Suitable micro-organisms for this type of study are the mycoplasmas, especially those closely cell-associated. Although the relationships between various mycoplasmas have been reported (Reich et al, 1966a, b ;Somerson et al, 1967;McGee et al, 1967), the work was not exhaustive and utilized either RNA-DNA hybridization or DNA-DNA hybridization in agar columns, which have limits of accuracy that impede the interpretation of low levels of hybridization.The present study utilized the more quantitative solution hybridization procedure followed by analysis of hybrid formation on hydroxyapatite columns (Britten & Kohne, 1966) in an attempt to provide a base of relationship data upon which to build a molecular detection system. In addition, this study utilized a large number of reciprocal hybridizations with 1251-labelled DNA allowing grouping of the micro-organisms into classes based on low degrees of homology.…”

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Partial Nucleotide Sequence Similarity within Species of Mycoplasma and Acholeplasma

1980

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333The nucleotide sequence relationships between 18 species of Mycoplasma and 3 species of Acholeplasma were examined by solution DNA hybridization. Radiolabelled DNAs from strains representing 13 Mycoplasma and 2 Acholeplasma species were used as hybridization probes, The mycoplasmas were heterogeneous but displayed a low degree of conserved information of the order of 3 to 5 % of the genome. However, several species within each genus showed 13 to 15 % homology. There was no detectable homology between species from the two genera, and M . pneumoniae and M . neurolyticum appeared to be unrelated to any of the other Mycoplasma species or to each other. These results suggest that it may be possible to isolate genes common to most, if not all, Mycoplasma and Acholeplasma species. I N T R O D U C T I O NThe use of nucleotide sequence homology as a taxonomic tool for bacteria and viruses is accepted practice. Likewise, measurement of nucleic acid hybridization kinetics has become a widely employed technique for detecting viruses in infected and transformed cells (Sharp et al., 1974). Before this technique can be extended to the detection of fastidious microorganisms in tissue a good understanding of the nucleotide sequence relationships within the microbial group is necessary. Suitable micro-organisms for this type of study are the mycoplasmas, especially those closely cell-associated. Although the relationships between various mycoplasmas have been reported (Reich et al., 1966a, b ;Somerson et al., 1967;McGee et al., 1967), the work was not exhaustive and utilized either RNA-DNA hybridization or DNA-DNA hybridization in agar columns, which have limits of accuracy that impede the interpretation of low levels of hybridization.The present study utilized the more quantitative solution hybridization procedure followed by analysis of hybrid formation on hydroxyapatite columns (Britten & Kohne, 1966) in an attempt to provide a base of relationship data upon which to build a molecular detection system. In addition, this study utilized a large number of reciprocal hybridizations with 1251-labelled DNA allowing grouping of the micro-organisms into classes based on low degrees of homology.This study demonstrates that there is little homology among different Mycoplasma and Acholeplasma species. However, there is a partial conservation of nucleotide sequences between several different species that are found in the same or similar hosts. M E T H O D SGrowth of mycoplasmas andpurifcation of DNA. with 10 % (v/v) whole horse serum (Irvine Scientific, Santa Ana, Calif., U.S.A.), 0.5 % (w/v) yeast extract and 100 units penicillin ml-l. Confirmation of growth and the absence of contamination were checked by plating 0.1 ml of the broth culture on PPLO agar supplemented as above. Before harvesting, cultures wereGram-stained to check for contamination. Uncontaminated cultures were centrifuged (40 min, 33 OO0 g) and the pellet was resuspended in 0.15 M-NaCl (1Oml) with sodium dodecyl sulphate (SDS, 1 %, w/v, h a 1 concn), 1 m-EDTA and 100...

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Molecular Genetic Studies of Relationships Among Mycoplasma, L‐forms and Bacteria*

Cited by 55 publications

References 24 publications

Bovine Mycoplasmas: Genome Size and Base Composition of DNA

Bovine Mycoplasmas: Genome Size and Base Composition of DNA

The antigens ofMycoplasma hominis

Partial Nucleotide Sequence Similarity within Species of Mycoplasma and Acholeplasma

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