scholarly journals Variable Surface Protein Vmm of Mycoplasma mycoides subsp. mycoides Small Colony Type

2002 ◽  
Vol 184 (13) ◽  
pp. 3712-3722 ◽  
Author(s):  
Anja Persson ◽  
Karin Jacobsson ◽  
Lars Frykberg ◽  
Karl-Erik Johansson ◽  
François Poumarat
Keyword(s):  
Phase Variation ◽  
Surface Protein ◽  
Transcriptional Level ◽  
Colony Type ◽  
Mycoplasma Capricolum ◽  
Binding Epitope ◽  
Variable Surface ◽  
Mycoplasma Mycoides ◽  
Small Colony ◽  
Reversible Phase

ABSTRACT A variable surface protein, Vmm, of the bovine pathogen Mycoplasma mycoides subsp. mycoides small colony type (M. mycoides SC) has been identified and characterized. Vmm was specific for the SC biotype and was expressed by 68 of 69 analyzed M. mycoides SC strains. The protein was found to undergo reversible phase variation at a frequency of 9 × 10−4 to 5 × 10−5 per cell per generation. The vmm gene was present in all of the 69 tested M. mycoides SC strains and encodes a lipoprotein precursor of 59 amino acids (aa), where the mature protein was predicted to be 36 aa and was anchored to the membrane by only the lipid moiety, as no transmembrane region could be identified. DNA sequencing of the vmm gene region from ON and OFF clones showed that the expression of Vmm was regulated at the transcriptional level by dinucleotide insertions or deletions in a repetitive region of the promoter spacer. Vmm-like genes were also found in four closely related mycoplasmas, Mycoplasma capricolum subsp. capricolum, M. capricolum subsp . capripneumoniae, Mycoplasma sp. bovine serogroup 7, and Mycoplasma putrefaciens. However, Vmm could not be detected in whole-cell lysates of these species, suggesting that the proteins encoded by the vmm-like genes lack the binding epitope for the monoclonal antibody used in this study or, alternatively, that the Vmm-like proteins were not expressed.

Characterization of the gene for an immunodominant 72 kDa lipoprotein of Mycoplasma mycoides subsp. mycoides small colony type

Microbiology ◽  
1996 ◽  
Vol 142 (12) ◽  
pp. 3515-3524 ◽  
Author(s):  
X. Cheng ◽  
J. Nicolet ◽  
R. Miserez ◽  
P. Kuhnert ◽  
M. Krampe ◽  
...  
Keyword(s):  
Colony Type ◽  
Mycoplasma Mycoides ◽  
Small Colony

scholarly journals Phylogeny of the Mycoplasma mycoides cluster based on analysis of five conserved protein-coding sequences and possible implications for the taxonomy of the group

2007 ◽  
Vol 57 (10) ◽  
pp. 2247-2258 ◽  
Author(s):  
Lucía Manso-Silván ◽  
Xavier Perrier ◽  
François Thiaucourt
Keyword(s):  
Housekeeping Genes ◽  
New Techniques ◽  
Protein Coding ◽  
Coding Sequences ◽  
Individual Gene ◽  
Mycoplasma Capricolum ◽  
Large Colony ◽  
Mycoplasma Mycoides ◽  
Small Colony ◽  
Distinct Branch

A phylogenetic tree of the Mycoplasma mycoides cluster was inferred from a set of concatenated sequences from five housekeeping genes (fusA, glpQ, gyrB, lepA and rpoB). The relevance of this phylogeny was reinforced by detailed analysis of the congruence of the phylogenies derived from each of the five individual gene sequences. Two subclusters were distinguished. The M. mycoides subcluster comprised M. mycoides subsp. mycoides biotypes Small Colony (SC) and Large Colony (LC) and M. mycoides subsp. capri. The latter two groups could not be clearly separated, which supports previous proposals that they be united into a single taxonomic entity. The Mycoplasma capricolum subcluster included M. capricolum subsp. capricolum, M. capricolum subsp. capripneumoniae and Mycoplasma sp. bovine group 7 of Leach, a group of strains that remains unassigned. This group constituted a distinct branch within this cluster, supporting its classification as a subspecies of M. capricolum. Mycoplasma cottewii and Mycoplasma yeatsii clustered in a group that was distinct from Mycoplasma putrefaciens and they were all clearly separated from the M. mycoides cluster. In conclusion, this approach has allowed us to assign phylogenetic positions to all members of the M. mycoides cluster and related species and has proved the need to adjust the existing taxonomy. Furthermore, this method may be used as a reference technique to assign an unequivocal position to any particular strain related to this cluster and may lead to the development of new techniques for rapid species identification.

scholarly journals Versatile Use of oriC Plasmids for Functional Genomics of Mycoplasma capricolum subsp. capricolum

2005 ◽  
Vol 71 (6) ◽  
pp. 2888-2893 ◽  
Author(s):  
Carole Janis ◽  
Carole Lartigue ◽  
Joachim Frey ◽  
Henri Wróblewski ◽  
François Thiaucourt ◽  
...  
Keyword(s):  
Homologous Recombination ◽  
Functional Genomics ◽  
Gene Inactivation ◽  
Heterologous Proteins ◽  
Gene Encoding ◽  
Colony Type ◽  
Genome Data ◽  
Mycoplasma Capricolum ◽  
Large Colony ◽  
Mycoplasma Mycoides

ABSTRACT Replicative oriC plasmids were recently developed for several mollicutes, including three Mycoplasma species belonging to the mycoides cluster that are responsible for bovine and caprine diseases: Mycoplasma mycoides subsp. mycoides small-colony type, Mycoplasma mycoides subsp. mycoides large-colony type, and Mycoplasma capricolum subsp. capricolum. In this study, oriC plasmids were evaluated in M. capricolum subsp. capricolum as genetic tools for (i) expression of heterologous proteins and (ii) gene inactivation by homologous recombination. The reporter gene lacZ, encoding β-galactosidase, and the gene encoding spiralin, an abundant surface lipoprotein of the related mollicute Spiroplasma citri, were successfully expressed. Functional Escherichia coli β-galactosidase was detected in transformed Mycoplasma capricolum subsp. capricolum cells despite noticeable codon usage differences. The expression of spiralin in M. capricolum subsp. capricolum was assessed by colony and Western blotting. Accessibility of this protein at the cell surface and its partition into the Triton X-114 detergent phase suggest a correct maturation of the spiralin precursor. The expression of a heterologous lipoprotein in a mycoplasma raises potentially interesting applications, e.g., the use of these bacteria as live vaccines. Targeted inactivation of gene lppA encoding lipoprotein A was achieved in M. capricolum subsp. capricolum with plasmids harboring a replication origin derived from S. citri. Our results suggest that the selection of the infrequent events of homologous recombination could be enhanced by the use of oriC plasmids derived from related mollicute species. Mycoplasma gene inactivation opens the way to functional genomics in a group of bacteria for which a large wealth of genome data are already available and steadily growing.

scholarly journals Phage display-based identification and potential diagnostic application of novel antigens from Mycoplasma mycoides subsp. mycoides small colony type

2010 ◽  
Vol 142 (3-4) ◽  
pp. 285-292 ◽  
Author(s):  
Shamoon Naseem ◽  
Jochen Meens ◽  
Joerg Jores ◽  
Martin Heller ◽  
Stefan Dübel ◽  
...  
Keyword(s):  
Phage Display ◽  
Colony Type ◽  
Diagnostic Application ◽  
Mycoplasma Mycoides ◽  
Small Colony

Humoral and bronchial immune responses in cattle experimentally infected with Mycoplasma mycoides subsp. mycoides small colony type

1998 ◽  
Vol 59 (2-3) ◽  
pp. 109-122 ◽  
Author(s):  
El-Mostafa Abdo ◽  
Jacques Nicolet ◽  
Raymond Miserez ◽  
Rosario Gonçalves ◽  
José Regalla ◽  
...  
Keyword(s):  
Immune Responses ◽  
Colony Type ◽  
Mycoplasma Mycoides ◽  
Small Colony

scholarly journals Distinctive Repertoire of Contingency Genes Conferring Mutation- Based Phase Variation and Combinatorial Expression of Surface Lipoproteins in Mycoplasma capricolum subsp. capricolum of the Mycoplasma mycoides Phylogenetic Cluster

2006 ◽  
Vol 188 (13) ◽  
pp. 4926-4941 ◽  
Author(s):  
Kim S. Wise ◽  
Mark F. Foecking ◽  
Kerstin Röske ◽  
Young Jin Lee ◽  
Young Moo Lee ◽  
...  
Keyword(s):  
Expression Patterns ◽  
Phase Variation ◽  
Wide Distribution ◽  
Surface Proteins ◽  
Dynamic Surface ◽  
Phylogenetic Cluster ◽  
Mycoplasma Capricolum ◽  
Gene Sets ◽  
Mycoplasma Mycoides ◽  
Stochastic Expression

ABSTRACT The generation of surface variation among many divergent species of Mollicutes (mycoplasmas) occurs through stochastic expression patterns of diverse lipoprotein genes. The size and wide distribution of such variable gene sets in minimal (∼0.6- to 1.4-Mb) mycoplasmal genomes suggest their key role in the adaptation and survival of these wall-less monoderms. Diversity through variable genes is less clearly established among phylogenetically similar mycoplasmas, such as the Mycoplasma mycoides cluster of ruminant pathogens, which vary widely in host range and pathobiology. Using (i) genome sequences from two members of this clade, Mycoplasma capricolum subsp. capricolum and M. mycoides subsp. mycoides small colony biotype (SC), (ii) antibodies to specific peptide determinants of predicted M. capricolum subsp. capricolum gene products, and (iii) analysis of the membrane-associated proteome of M. capricolum subsp. capricolum, a novel set of six genes (vmcA to vmcF) expressing distinct Vmc (variable M. capricolum subsp. capricolum) lipoproteins is demonstrated. These occur at two separate loci in the M. capricolum subsp. capricolum genome, which shares striking overall similarity and gene synteny with the M. mycoides subsp. mycoides SC genome. Collectively, Vmc expression is noncoordinate and combinatorial, subject to a single-unit insertion/deletion in a 5′ flanking dinucleotide repeat that governs expression of each vmc gene. All vmc genes share modular regions affecting expression and membrane translocation. In contrast, vmcA to vmcD genes at one locus express surface proteins with highly structured size-variable repeating domains, whereas vmcE to vmcF genes express products with short repeats devoid of predicted structure. These genes confer a distinctive, dynamic surface architecture that may represent adaptive differences within this important group of pathogens as well as exploitable diagnostic targets.

scholarly journals Flow Cytometric Determination of the Effects of Antibacterial Agents on Mycoplasma agalactiae, Mycoplasma putrefaciens, Mycoplasma capricolum subsp. capricolum, and Mycoplasma mycoides subsp. mycoides Large Colony Type

2006 ◽  
Vol 50 (8) ◽  
pp. 2845-2849 ◽  
Author(s):  
Patricia Assunção ◽  
Nuno T. Antunes ◽  
Ruben S. Rosales ◽  
Carlos Poveda ◽  
Jose B. Poveda ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Propidium Iodide ◽  
Antibacterial Agents ◽  
Mycoplasma Species ◽  
Colony Type ◽  
Flow Cytometric ◽  
Mycoplasma Capricolum ◽  
Large Colony ◽  
Mycoplasma Mycoides

ABSTRACT Flow cytometry together with SYBR green I and propidium iodide was used to study the effects of enrofloxacin, ciprofloxacin, gentamicin, chloramphenicol, oxytetracycline, and tylosin on four mycoplasma species. Inhibition of mycoplasma growth could be detected by as early as 3 h after the start of treatment. The strongest effect was observed with enrofloxacin- and ciprofloxacin-treated cells.

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