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Query: UNIPROT:P06889 (Mol)
 630,302 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Recently, a 16S rDNA-based polymerase chain reaction (PCR) assay was developed for the selective and sensitive detection of Mycoplasma pirum. In this study, the same procedure was used in order to selectively detect by PCR two human mycoplasmas, M. hominis and M. penetrans, with a high level of sensitivity even in a context of human DNA. For each assay, the specificity was verified by testing DNA from other mollicute species (including those closely related to the corresponding mycoplasma), from bacteria phylogenetically close to mollicutes, from Escherichia coli and from human peripheral blood mononuclear cells (PBMCs). Each assay proved to be highly sensitive since it reliably detected 10 DNA molecules, even in a context of human DNA. The results of this study demonstrate the suitability of our procedure using primers which were designed for the PCR detection of human mollicutes with a high specificity and a low and reproducible threshold of sensitivity.
Mol Cell Probes 1994 Apr
PMID:Development of PCR-based assays for the detection of two human mollicute species, Mycoplasma penetrans and M. hominis. 793 12

Eleven hitherto unknown Mycoplasma pneumoniae proteins were identified and characterized with respect to their size and subcellular location. This was carried out through the construction of in vitro gene fusions between a modified mouse dehydrofolate reductase (dhfr) gene and selected regions (cosmid clones) of the M. pneumoniae genome and expressing them in Escherichia coli. Positive clones were identified using antibodies against specific fractions of M. pneumoniae. The deduced protein sequences of 11 out of 30 clones did not show significant homologies to known proteins in protein data-bank searches. Monospecific antibodies against these 11 fusion proteins were used to determine the size and cellular location of the corresponding M. pneumoniae proteins by immunoscreening Western blots of SDS-acrylamide gels from M. pneumoniae cell extracts.
Mol Microbiol 1994 Jul
PMID:Identification and characterization of hitherto unknown Mycoplasma pneumoniae proteins. 798 11

Insertion sequence (IS) elements are mobile genetic elements found in prokaryotes. We have identified a repetitive element from Mycoplasma pulmonis, a murine pathogen, that is similar to eubacterial IS elements. By subcloning a single strain of M. pulmonis, we isolated a variant clone in which the IS element had undergone an apparent transposition event. The nucleotide sequences of the element, designated IS1138, and the target site into which it inserted were determined. IS1138 consists of 1288 bp with 18 bp perfect terminal inverted repeats. Sequence analysis of the target site before and after insertion of IS1138 identified a 3 bp duplication of target DNA flanking the element. The predicted amino acids encoded by the major open reading frame of IS1138 share significant similarity with the transposases of the IS3 family. Southern hybridization analysis indicates that repetitive sequences similar to IS1138 are present in most, if not all, strains of M. pulmonis, but IS1138-like sequences were not detected in other mycoplasmal species.
Mol Microbiol 1993 Feb
PMID:Identification and characterization of IS1138, a transposable element from Mycoplasma pulmonis that belongs to the IS3 family. 809 21

Mycoplasma hyopneumoniae (Mhp) is the etiologic agent of mycoplasma pneumonia in swine. The purpose of this study was to develop a species-specific polymerase chain reaction (PCR)-based diagnostic reagent for the identification of Mhp. Based upon DNA sequence analysis of a cloned fragment of Mhp DNA, PCR primers were constructed and tested against different strains of Mhp, Mycoplasma flocculare, other mycoplasma species, and non-Mollicute micro-organisms which commonly inhabit the respiratory tracts of swine. A total of 40 field isolates from Mhp and four field isolates of M. flocculare have been examined. Positive signals were obtained in PCR with Mhp reference strains and all 40 Mhp field isolates, but not with other Mollicutes micro-organisms.
Mol Cell Probes 1993 Oct
PMID:Development of polymerase chain reaction primers to detect Mycoplasma hyopneumoniae. 826 72

Mycoplasma pneumoniae-specific monoclonal antibodies were constructed for the purpose of developing reagents for clinical diagnostics. The monoclonal antibodies, designated Mp2B12 and Mp3A11, recognize surface exposed antigens of 28 kDa and 170 kDa, respectively. Mp3A11 recognizes an epitope on a cytadhesin molecule, P1, which is not shared on the analogous cytadhesin of M. genitalium. Both monoclonal antibodies are capable of detecting 1 x 10(5) M. pneumoniae in a standard ELISA test.
Mol Cell Probes 1993 Apr
PMID:Development of monoclonal antibodies for the detection of Mycoplasma pneumoniae. 832 Dec 52

A polymerase chain reaction (PCR) system was developed for the detection of mollicutes as contaminants of cell cultures. By using three oligonucleotides chosen in the 16S rDNA sequences, two sets of primers able to promote amplification of all Mycoplasma and Ureaplasma (molli1-molli2a) or all Acholeplasma (molli1-molli2b) species examined were determined. This PCR system, first applied to experimentally infected Vero cell lines, was then evaluated for the detection of mollicutes in 86 cell culture samples, comparatively to DNA staining, culture and ELISA. The results obtained by the four techniques were in agreement in 82 cases (36 positive, 46 negative). PCR allowed detection of contamination in one and two cases negative by ELISA and culture, respectively, and confirmed questionable results obtained by DNA staining. As described, PCR seems to be a very convenient tool for routine detection of cell culture contaminants.
Mol Cell Probes 1993 Jun
PMID:Detection of mollicute contamination in cell cultures by 16S rDNA amplification. 836 66

Fibroblasts may play an important role in the modulation of immune and inflammatory responses through elaboration of cytokines. To test this hypothesis, human lung fibroblasts were isolated from transbronchial biopsy specimens and assayed for production of interleukin-6 (IL-6) and granulocyte/macrophage colony-stimulating factor (GM-CSF). The sources of fibroblasts included lung allografts, recipient lungs obtained at time of transplant, and normal lung tissue removed during tumor resection. During the course of these studies, several early-passage fibroblasts from transplant recipients were observed to contain mycoplasma (MP)-like organisms as detected by extranuclear fluorescent staining with Hoechst 33258. Positive staining cultures were associated with isolation of Mycoplasma fermentans. IL-6 and GM-GSF as measured by ELISA were found to be elevated over 50-fold in conditioned medium from MP-infected fibroblasts as compared with noninfected lines. Treatment of cells with mycoplasma removal agent (MRA) eliminated extranuclear Hoechst fluorescence and significantly reduced the production of these cytokines. Tumor necrosis factor-beta (TNF-beta) induction of IL-6 and GM-CSF was amplified synergistically in infected cultures. No additional production of IL-6 or GM-CSF was observed in infected cultures treated with interferon-gamma (IFN-gamma) despite the ability of IFN-gamma to modestly induce IL-6 in uninfected cultures. Thus, in vitro infection of lung fibroblasts with MP represents a potent stimulus for the production of inflammatory cytokines and, therefore, necessitates rigorous control for these organisms in cell culture studies.(ABSTRACT TRUNCATED AT 250 WORDS)
Am J Respir Cell Mol Biol 1993 Apr
PMID:Enhanced secretion of immune-modulating cytokines by human lung fibroblasts during in vitro infection with Mycoplasma fermentans. 847 29

To investigate the reading properties of adenosine in the wobble position we have used site-directed mutagenesis of the Escherichia coli glycine tRNA1(CCC) gene to substitute the nucleotide A in the wobble position of the corresponding tRNA. The effect of this change on the ability of the tRNA to discriminate between the nucleotides in the third position of the glycine codons has been investigated. We have compared the ability of the mutant glycine tRNA1(UCC) and glycine tRNA1(ACC) as well as the mycoplasma glycine tRNA(UCC) to read the glycine codons. The results showed that glycine tRNA1(ACC) unlike glycine tRNA1(UCC) did not fully discriminate between the glycine codons. These experiments were carried out using a new in vitro protein synthesizing system that allows us to monitor the reading of all four glycine codons. In the present paper we give a detailed description of this new in vitro system.
J Mol Biol 1993 Apr 05
PMID:Undiscriminating codon reading with adenosine in the wobble position. 847 31

Test systems for rapid detection of mycoplasmas in biological samples have been elaborated on the base of the polymerase chain reaction (PCR). Amplification of the conservative rDNA sequences was used for testing of cell cultures for mycoplasmal contamination. Mycoplasma pneumoniae detection system has been developed based on amplification of the species-specific DNA sequences. Inversions of some repeated sequences in the Mycoplasma pneumoniae genome make it possible to run the PCR with a single primer. The revealed spacer length polymorphism for 16S-23S rDNA operons can be used in the mycoplasmas identification.
Mol Gen Mikrobiol Virusol
PMID:[Diagnosis of mycoplasma infections using directed amplification]. 851 51

We have used the polymerase chain reaction (PCR) to detect Mycoplasma hyopneumoniae in tracheobronchiolar washings collected from experimentally infected piglets. On the basis of the published nucleotide sequence of M. hyopneumoniae I141 probe (accession number U02537), primers were chosen to produce an amplified fragment of 1561 bp. All the M. hyopneumoniae strains tested could be detected by the PCR test. DNA from other mycoplasmal and bacterial species currently isolated from respiratory tract of piglets gave negative result. The detection limit was estimated to be 500 fg of purified DNA corresponding to 4.10(2) organisms. The sensitivity of PCR reaction was also evaluated on microorganisms in culture, the limit sensitivity was 2.5 10(3) organisms. In the present study, a total of 143 tracheobronchiolar washings collected from experimentally infected piglets were submitted to PCR. For each tracheobronchiolar washing, PCR was performed on crude extracts treated with lysis buffer and on extracted DNA. The PCR results obtained with the two kinds of samples were compared to the immunofluorescence (IF) results. This comparison indicates a good correlation between PCR and IF test in 121/143 cases. The presence of M. hyopneumoniae is revealed in 19/143 of the washing samples only by PCR. In our hand, PCR appears to be the more sensitive test to detect M. hyopneumoniae in experimentally infected piglets.
Mol Cell Probes 1996 Feb
PMID:Polymerase chain reaction for Mycoplasma hyopneumoniae detection in tracheobronchiolar washings from pigs. 868 72


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