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.
Department of Microbiology and Immunology, 209 Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill 27599-7295, USA.
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D001483:Base Sequence; D015152:Blotting, Northern; D018920:Coculture Techniques; D017931:DNA Primers; D015342:DNA Probes / Q000737:chemistry; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D020869:Gene Expression Profiling; D005798:Genes, Bacterial; D006801:Humans; D008969:Molecular Sequence Data; D009174:Mycoplasma / Q000737:chemistry / Q000235:genetics*; D012329:RNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D014407:Tumor Cells, Cultured
D017931:DNA Primers; D015342:DNA Probes; D004269:DNA, Bacterial; D012329:RNA, Bacterial