Identification of novel Mycoplasma hyorhinis gene fragments by differential display analysis of co-cultures.

Mycoplasmas are a diverse group of wall-less prokaryotes that have evolved an unusually small genome by adopting a parasitic mode of life. Recently, intense efforts have been made to sequence mycoplasma genomes and to define a minimal genome using mycoplasma as a model. Due to their parasitic nature, mycoplasma species are often difficult to cultivate, making it challenging to identify and sequence mycoplasma genes. In this report, we describe a method for identifying mycoplasma gene fragments from co-cultures using differential display analysis. Using this technique, we have identified fragments of seven putative genes from Mycoplasma hyorhinis. Sequence similarities suggest that four of these genes are members of the proposed minimal mycoplasma genome. The application of differential display analysis to co-cultures should be useful in the identification of genes from a variety of pathogenic organisms that are difficult to cultivate without a host.

[1]  A F Angulo,et al.  Genus- and species-specific identification of mycoplasmas by 16S rRNA amplification , 1992, Applied and environmental microbiology.

[2]  Manuel Peitsch,et al.  A genome-based approach for the identification of essential bacterial genes , 1998, Nature Biotechnology.

[3]  Thomas L. Madden,et al.  Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. , 1997, Nucleic acids research.

[4]  J. Ting,et al.  Identification of Class II Transcriptional Activator-Induced Genes by Representational Difference Analysis: Discoordinate Regulation of the DNα/DOβ Heterodimer1 , 2000, The Journal of Immunology.

[5]  E. Koonin,et al.  A minimal gene set for cellular life derived by comparison of complete bacterial genomes. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[6]  R. Gardella,et al.  Growth of Mycoplasma hyorhinis cultivar alpha on semisynthetic medium , 1995, Applied and environmental microbiology.

[7]  T. Umemura,et al.  Demonstration of Mycoplasma hyorhinis as a Possible Primary Pathogen for Porcine Otitis Media , 1995, Veterinary pathology.

[8]  A. Miettinen,et al.  Mycoplasmas and arthritis. , 1983, Zeitschrift fur Rheumatologie.

[9]  R. Parish,et al.  A mycoplasmal protein influences tumour cell invasiveness and contact inhibition in vitro. , 1990, Journal of cell science.

[10]  S. Razin Adherence of Pathogenic Mycoplasmas to Host Cells , 1999, Bioscience reports.

[11]  T. Chen,et al.  In situ detection of mycoplasma contamination in cell cultures by fluorescent Hoechst 33258 stain. , 1977, Experimental cell research.

[12]  Shmuel Razin,et al.  Molecular Biology and Pathogenicity of Mycoplasmas , 1998, Microbiology and Molecular Biology Reviews.

[13]  M. Kurimoto,et al.  Metastasis‐Promoting Activity of a Novel Molecule, Ag 243‐5, Derived from Mycoplasma, and the Complete Nucleotide Sequence , 1995, Microbiology and immunology.

[14]  R. Fleischmann,et al.  The Minimal Gene Complement of Mycoplasma genitalium , 1995, Science.

[15]  B. Mallard,et al.  Mycoplasma hyorhinis infection of pigs selectively bred for high and low immune response. , 1998, Veterinary immunology and immunopathology.

[16]  L. R. Finch,et al.  Mycoplasmas: molecular biology and pathogenesis. , 1992 .

[17]  G. Stark,et al.  Mutant human cells defective in induction of major histocompatibility complex class II genes by interferon gamma. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[18]  C. Stanners,et al.  Identification of a Human Cancer Related Organ‐Specific Neoantigen , 1993, Microbiology and immunology.

[19]  R. A. Del Giudice,et al.  Cell culture models as ancillary tools in the isolation and characterization of mycoplasmas. , 1984, Israel journal of medical sciences.

[20]  H. Hilbert,et al.  Complete sequence analysis of the genome of the bacterium Mycoplasma pneumoniae. , 1996, Nucleic acids research.

[21]  B. Mallard,et al.  Cytokines in Mycoplasma hyorhinis-Induced Arthritis in Pigs Bred Selectively for High and Low Immune Responses , 2000, Infection and Immunity.

[22]  O. White,et al.  Global transposon mutagenesis and a minimal Mycoplasma genome. , 1999, Science.

[23]  T. Umemura,et al.  Intranasally Inoculated Mycoplasma hyorhinis Causes Eustachitis in Pigs , 1999, Veterinary pathology.