Cloned genomic DNA sequences from Mycoplasma hyorhinis encoding antigens expressed in Escherichia coli.

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Abstract
A library of cloned M. hyorhinis genomic sequences was constructed by incorporation of EcoRI digestion fragments of mycoplasma DNA into the .lambda. Charon 4A phage vector. Immunological screening of recombinant phage plaques identified clones containing genes encoding mycoplasma antigenic structures expressed in an E. coli host. Two such recombinant phage isolates, .lambda.Ch4A-MhrG1 and .lambda.Ch4A-MhrG28, were defined and contain distinct genomic sequences by analysis of restriction endonuclease fragments. Inoculation of mice with recombinant gene products from .lambda.Ch4A-MhrG1 yielded antiserum selectively recognizing a MW 29,500 trypsin-sensitive mycoplasma constituent. This established a means for producing selected immunogenic mycoplasma components in a bacterial host. The cloned genomic sequences of M. hyorhinis encoding expressed mycoplasma antigens represent molecular probes that can be characterized both by specific DNA sequences and by the antigenic structure of corresponding gene products. These genomic fragments define initial physical markers of the M. hyorhinis genome and may be useful in assessing antigenic and molecular genetic relationships within the genus Mycoplasma and among other members of the class Mollicutes.