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We report the construction of a physical map of the genome of the human pathogen Mycoplasma genitalium through the use of pulse-field gel electrophoresis. The small size and relative simplicity of this genome permit the arrangement of restriction fragments without having to construct linking clones. The size of the genome has been calculated to be approximately 600 kb and several important genetic determinants have been assigned specific loci on the map.
Mol Microbiol 1990 Apr
PMID:A physical map of the Mycoplasma genitalium genome. 235 75

Genomic restriction fragments isolated from Mycoplasma hyorhinis and Mycoplasma hyopneumoniae were shown by DNA hybridization and nucleotide sequence analyses to contain sequences common to these two species, as well as another porcine-derived mycoplasma, Mycoplasma flocculare. Intraspecies hybridization experiments using these fragments as probes indicated that the sequence is highly redundant in several strains of M. hyorhinis, but that there is diversity in the sizes of restriction fragments detected among these strains. In contrast, repetition of the sequence was limited in M. hyopneumoniae and M. flocculare, and no homologies to this repeated element were apparent in mycoplasma species isolated from animal hosts other than the swine. The reiterated sequence may reflect intraspecies genomic diversification in M. hyorhinis and its selective presence in otherwise unrelated species raises the possibility that it has been horizontally transmitted between these organisms.
Mol Microbiol 1988 Sep
PMID:Reiterated DNA sequences defining genomic diversity within the species Mycoplasma hyorhinis. 246 Jul 18

The nucleotide sequences of the complete set of tRNA species in Mycoplasma capricolum, a derivative of Gram-positive eubacteria, have been determined. This bacterium represents the first genetic system in which the sequences of all the tRNA species have been determined at the RNA level. There are 29 tRNA species: three for Leu, two each for Arg, Ile, Lys, Met, Ser, Thr and Trp, and one each for the other 12 amino acids as judged from aminoacylation and the anticodon nucleotide sequences. The number of tRNA species is the smallest among all known genetic systems except for mitochondria. The tRNA anticodon sequences have revealed several features characteristic of M. capricolum. (1) There is only one tRNA species each for Ala, Gly, Leu, Pro, Ser and Val family boxes (4-codon boxes), and these tRNAs all have an unmodified U residue at the first position of the anticodon. (2) There are two tRNAThr species having anticodons UGU and AGU; the first positions of these anticodons are unmodified. (3) There is only one tRNA with anticodon ICG in the Arg family box (CGN); this tRNA can translate codons CGU, CGC and CGA. No tRNA capable of translating codon CGG has been detected, suggesting that CGG is an unassigned codon in this bacterium. (4) A tRNATrp with anticodon UCA is present, and reads codon UGA as Trp. On the basis of these and other observations, novel codon recognition patterns in M. capricolum are proposed. A comparatively small total, 13, of modified nucleosides is contained in all M. capricolum tRNAs. The 5' end nucleoside of the T psi C-loop (position 54) of all tRNAs is uridine, not modified to ribothymidine. The anticodon composition, and hence codon recognition patterns, of M. capricolum tRNAs resemble those of mitochondrial tRNAs.
J Mol Biol 1989 Sep 05
PMID:Codon recognition patterns as deduced from sequences of the complete set of transfer RNA species in Mycoplasma capricolum. Resemblance to mitochondria. 247 13

The genetic code, once thought to be "frozen," shows variations from the universal code. Variations are found in mitochondria, Mycoplasma, and ciliated protozoa. The variations result from reassignment of codons, especially stop codons. The reassignments take place by disappearance of a codon from coding sequences, followed by its reappearance in a new role. Simultaneously, a changed anticodon must appear. We discuss the role of directional mutation pressure in the events, and we also describe the possibility that such events have taken place during early evolution of the genetic code and can occur during its present evolution.
J Mol Evol 1989 Apr
PMID:Codon reassignment (codon capture) in evolution. 249 83

The sequences of Saccharomyces carlsbergensis ribosomal protein (r-protein) SL25* and its equivalents from Candida utilis (CL25), Escherichia coli (EL23), Bacillus stearothermophilus (BL23), Mycoplasma capricolum (ML23), Marchantia polymorpha chloroplasts (McpL23), and Nicotiana tabacum chloroplasts (NcpL23) were examined using a computer program that evaluates the extent of sequence similarity by calculating correlation coefficients for each pair of residues in two proteins from a number of physical properties of individual amino acids. Comparison matrices demonstrate that the prokaryotic sequences (including McpL23 and NcpL23) can be aligned unambiguously by introducing small internal deletions/insertions at three specific positions. A similar comparison brought to light a clear evolutionary relationship between the prokaryotic and the yeast proteins despite the fact that visual inspection of these sequences revealed only limited similarity. The alignment deduced from this comparison shows the two yeast r-proteins to have acquired a long (50-60 amino acids) N-terminal extension as well as a 13-amino acid-long deletion near the C-terminus. The significance of these findings in terms of the evolution of r-proteins in general and the biological function of various parts of the SL25 protein in particular is discussed.
J Mol Evol 1989 May
PMID:Structural comparison of 26S rRNA-binding ribosomal protein L25 from two different yeast strains and the equivalent proteins from three eubacteria and two chloroplasts. 250 3

Nucleotide sequence analysis of two Mycoplasma hyopneumoniae-derived copies of a repetitive genetic element revealed structural similarities to typical prokaryotic insertion sequences. This is the first such sequence identified in the class Mollicutes. The element spans approximately 1550bp, with 28bp inverted terminal repeats. Two open reading frames occur within the sequence, one potentially encoding a protein with a size-variant alpha-helical domain containing heptameric leucine periodicity. Hybridization data with several strains from each of two mycoplasma species showed that the repetitive sequence is variably distributed within the M. hyopneumoniae and Mycoplasma hyorhinis chromosomes and indicated that in some cases the repeated sequence is contained within a larger genetic element which may be the result of phage or plasmid insertion.
Mol Microbiol 1989 Jul
PMID:A Mycoplasma genetic element resembling prokaryotic insertion sequences. 255 57

In this study we performed a cytochemical comparison of peroxidase and acid phosphatase activities in peritoneal eosinophils from specific pathogen-free (SPF) and Mycoplasma pulmonis-infected rats. When eosinophils ingested polystyrene particles for 120 min, peroxidase- and acid phosphatase-positive specific granules, as well as small granules, fused to phagosomes. Unusual peroxidase activity was detected in some particle-containing phagosomes in 6.8% of eosinophils from control SPF rats and 31% of those from infected rats. Intense acid phosphatase activity was also demonstrated in some phagosomes in 4 and 20% of eosinophils from control and infected rats, respectively. The rate of peroxidase-positive and acid phosphatase-positive phagosomes to all ingested phagosomes was 8.1 and 7.3% in control rats and rose to 41.3 and 31.1% in those infected, respectively. The population of eosinophils (18%) in the control peritoneal cells did not change after infection (16.6%). These results suggest that the intraphagosomal release of lysosomal enzymes was significantly stimulated in peritoneal eosinophils of the rats spontaneously infected with M. pulmonis.
Exp Mol Pathol 1989 Dec
PMID:Stimulated intraphagosomal release of eosinophil peroxidase and acid phosphatase in the rats spontaneously infected with Mycoplasma pulmonis: a cytochemical study. 259 62

Enzootic porcine pneumonia is caused by Mycoplasma hyopneumoniae. Since the disease is of world-wide importance it is important to detect and identify the causative agent. In experience laboratories this mycoplasma can usually be detected by culture but its identification still is difficult and time consuming. We have cloned random Eco R1 fragments of M. hyopneumoniae DNA to M13mp19 and used the resultant recombinant to produce a probe capable of detecting approximately 10 pg of the mycoplasma DNA (10(4) organisms). By using appropriate stringency the test was made specific for M. hyopneumoniae, although at lower stringency reaction was positive with Mycoplasma flocculare at 1000 x the concentration limit. The assay did not detect M. hyopneumoniae in DNA from lungs of chronically infected animals but it did react with DNA isolated from the organisms cultured from the infected lung material.
Mol Cell Probes 1989 Sep
PMID:A gene probe to detect Mycoplasma hyopneumoniae, the etiological agent of enzootic porcine pneumonia. 279 7

A 5.5 kilobase DNA fragment from an Eco RI digest of the Mycoplasma gallisepticum genome was specific for the detection of M. gallisepticum. This 5.5 kb fragment was initially cloned into bacteriophage lambda gt11 followed by subcloning into the plasmid vector pGEM-3Z. The incorporation of a biotin label was accomplished by utilizing biotin-11-dUTP in a nick translation reaction. This probe, designated pMg6, reacts specifically with M. gallisepticum when tested against various mycoplasma DNAs in Southern blot hybridization analysis. Spot-blot hybridization data indicate the pMg6 is capable of detecting 800 pg of M. gallisepticum DNA.
Mol Cell Probes 1988 Sep
PMID:Species-specific biotinylated DNA probe for the detection of Mycoplasma gallisepticum. 322 85

Mycoplasma capricolum uses two tryptophan codons, the "universal" nonsense codon UGA and the universal codon UGG. The bacterium contains two tryptophan tRNAs, one with anticodon UCA, (U: 2'-O-methyl U derivative), and the other with CCA (5'-C: partially 2'-O-methylated). tRNAUCA would translate codons UGA and probably UGG by wobbling. tRNACCA is much less charged by tryptophan in the cells than tRNAUCA, and the intracellular amount of tRNACCA is 5-10 times lower than that of tRNAUCA. The genes for these two tRNAs are separated by a terminator-like structure in a single operon. In vitro transcription experiments suggest that the predominance of tRNAUCA over tRNACCA results from the attenuation of transcription by this terminator-like structure.
Mol Gen Genet 1988 May
PMID:Evolutionary dynamics of tryptophan tRNAs in Mycoplasma capricolum. 340 3


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