Molecular biology and genetics of mycoplasmas (Mollicutes)

Razin, Shmuel

doi:10.1128/mr.49.4.419-455.1985

Cited by 158 publications

(14 citation statements)

References 182 publications

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“…The CGC values inferred, as above, from the GGC value for this gene were within a range of 5 mol% from the RGC for 100 prokaryotic strains that had not been previously included in the determination of correlation. Furthermore, we observed that the CGC values obtained for such prokaryotic species as Campylobacter jejuni (31?0 and 31?2 mol%), Ureaplasma urealyticum (25?9 mol%) or T. whipplei (49?3 mol%) were closer to their RGC [30?3 and 30?5 (Owen, 1983), 25?5 (Razin, 1985) and 46?3 mol% , respectively] than the DNA G+C content values obtained using traditional methods [31?5 (Owen, 1983), 26?9-28 (Razin, 1985 and 59?4 mol% (La Scola et al, 2001), respectively].…”

Section: Cog Numbermentioning

confidence: 96%

Estimation of prokaryote genomic DNA G+C content by sequencing universally conserved genes

Fournier

Suhre

Fournous

et al. 2006

International Journal of Systematic and Evolutionary Microbiology

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Determination of the DNA G+C content of prokaryotic genomes using traditional methods is time-consuming and results may vary from laboratory to laboratory, depending on the technique used. We explored the possibility of extrapolating the genomic DNA G+C content of prokaryotes from gene sequences. For this, 127 universally conserved genes were studied from 50 prokaryotic genomes in the Clusters of Orthologous Groups database. Of these, 57 genes were present as a single copy in the genomes of 157 different prokaryote species available in GenBank. There was a strong correlation [coefficient of determination (r 2 ) >95 %] between the DNA G+C contents of 20 genes and their corresponding genomes. For each of the 157 prokaryotic genomes studied, the DNA G+C content of the 20 genes was used to determine a 'calculated' genome DNA G+C content (CGC) and this value was compared with the 'real' genome DNA G+C content (RGC). In order to select the most suitable gene for the determination of CGC values, we compared the r 2 and median mol% difference between CGC and RGC as well as the sensitivity of each gene to provide CGC values for prokaryotic genomes that differ by less than 5 mol% from their RGC. The highly conserved ftsY gene (median size 1144 nucleotides), a vertically inherited member of the GTPase superfamily, showed the highest r 2 value of 0?98, the smallest median mol% difference between CGC and RGC of 1?06 and a sensitivity of 100 %. Using ftsY DNA G+C content values, the CGC values of 100 genomes not included in the calculation of r 2 differed by less than 5 mol% from their RGC values. These data suggest that the genomic DNA G+C content of prokaryotes may be estimated easily and reliably from the ftsY gene sequence. INTRODUCTIONThe current taxonomic classification of prokaryotes is based on polyphasic taxonomy (Vandamme et al., 1996). This approach combines the genomic and phenotypic characteristics of a strain. The minimum amount of genomic information required for the description of a novel bacterial species must include its phylogenetic classification, DNA-DNA relatedness and the mol% G+C content of DNA (Stackebrandt et al., 2002). Previously, it has been suggested that micro-organisms showing more than 10 mol% difference in DNA G+C contents might not belong to the same genus and that 5 mol% is the common range found within a species (Goodfellow et al., 1997). Of the various methods available for the determination of DNA G+C content (De Ley, 1970;Ko et al., 1977;Mesbah & Whitman, 1989;Owen et al., 1969;Schildkraut et al., 1962;Xu et al., 2000), the thermal denaturation temperature (T m ) method is most commonly used . However, thermal denaturation is a time-consuming method that requires a large amount of DNA and lacks intra-and inter-laboratory reproducibility, and the T m is calculated using a formula proposed by Mandel et al. (1970) which is not suitable for prokaryotes with very low or elevated DNA G+C contents (Ezaki et al., 1990).In our laboratory, we have been studying the rpoB gene, encoding the b-subun...

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Section: Cog Numbermentioning

confidence: 96%

Estimation of prokaryote genomic DNA G+C content by sequencing universally conserved genes

Fournier

Suhre

Fournous

et al. 2006

International Journal of Systematic and Evolutionary Microbiology

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“…The closest relatives of H. felis , including Haemobartonella muris , Eperythrozoon suis , and Eperythrozoon wenyonii , are members of the order Mycoplasmatales, not Rickettsiales, on the basis of sequence analysis of the 16S rRNA gene 7,21,23,26. Mycoplasmas are known to carry only one or two copies of the 16S rRNA gene;25 therefore, conventional ISH should not detect the genomic 16S rRNA sequence in these species, and additional techniques are needed to enhance the sensitivity of the assay.…”

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Specific In Situ Hybridization of Haemobartonella felis with a DNA Probe and Tyramide Signal Amplification

et al. 2000

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Haemobartonella felis is an epierythrocytic bacterium suspected to be the causative agent of feline infectious anemia. Previous studies with a polymerase chain reaction assay have identified a mycoplasmal 16S rRNA gene sequence that coincides with clinical disease and the presence of organisms in the blood. Tissues from a cat experimentally infected with H. felis were used for in situ hybridization studies to physically link this 16S rRNA gene to the organisms on the red cells. A biotin-labeled probe was used in conjunction with tyramide signal amplification to visualize the hybridization signal. This study clearly demonstrates a specific hybridization signal on the red cells in the tissues of the H. felis-infected cat. This in situ hybridization study is the final step in fulfilling the molecular guidelines for disease causation and proves that H. felis, a mycoplasmal organism, is the causative agent of feline infectious anemia.

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“…They possess the smallest genome of any free-living organism, and the GϩC content of their DNA is among the lowest known. 21,22 The pig is host to a number of mycoplasmas, and the most commonly isolated species are Mycoplasma hyopneumoniae, M. hyorhinis, M. hyosynoviae, and M. flocculare. 23 Enzootic pneumonia caused by M. hyopneumoniae is a contagious respiratory disease characterized by chronic cough, growth retardation, low mortality, and high morbidity.…”

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confidence: 99%

Detection and Localization ofMycoplasma hyopneumoniaeDNA in Lungs from Naturally Infected Pigs by In Situ Hybridization Using a Digoxigenin-labeled Probe

Kwon

Chae

1999

Vet Pathol

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Mycoplasma hyopneumoniae DNA was detected in 20 naturally infected pigs by in situ hybridization using a nonradioactive digoxigenin-labeled DNA probe. A 520-base-pair DNA probe targeting a reiterative sequence of the M. hyopneumoniae genome was generated by the polymerase chain reaction. All 20 pigs infected with M. hyopneumoniae had distinct and positive hybridization signals without background staining. A strong hybridization signal was detected mainly in the luminal surface of bronchial and bronchiolar lining epithelial cells, whereas no hybridization signal was seen in the cytoplasm of bronchial and bronchiolar lining epithelial cells. When hybridization signal was detected in the luminal surface of bronchial and bronchiolar lining epithelial cells, a given bronchus or bronchiole had peribronchiolar lymphoid hyperplastic tissues. Hybridization signals were not seen in the peribronchiolar lymphoid hyperplastic tissues. A less intense signal was detected in the interstitial and alveolar macrophages randomly scattered in the thickened alveolar septa and spaces. Hybridization signal was rarely detected in the type I pneumocytes. The in situ hybridization technique developed in this study was useful for detection of M. hyopneumoniae nucleic acids in tissues taken from naturally infected piglets and may be a valuable technique for studying the pathogenesis of M. hyopneumoniae infection.

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Molecular biology and genetics of mycoplasmas (Mollicutes)

Cited by 158 publications

References 182 publications

Estimation of prokaryote genomic DNA G+C content by sequencing universally conserved genes

Estimation of prokaryote genomic DNA G+C content by sequencing universally conserved genes

Specific In Situ Hybridization of Haemobartonella felis with a DNA Probe and Tyramide Signal Amplification

Detection and Localization ofMycoplasma hyopneumoniaeDNA in Lungs from Naturally Infected Pigs by In Situ Hybridization Using a Digoxigenin-labeled Probe

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