Cloning of Mycoplasma pneumoniae DNA and expression of P1-epitopes in Escherichia coli.
A genomic library of Mycoplasma pneumoniae was constructed by cloning partial Sau3A-digested genomic DNA into the expression plasmids pEX1 to pEX3. The recombinant clones were screened for production of M. pneumoniae P1-antigen by an in situ colony enzyme-linked immunosorbent assay (ELISA) blot method with a monospecific rabbit antiserum raised against the surface protein P1. The length of the translated P1-sequence and the size of the inserted DNA were determined. By comparison it was shown that six clones contained DNA fragments coding for an internal part of the P1-protein and eight clones code for the C-terminal part of terminated P1-protein. In reactions against sera from patients suffering from M. pneumoniae infection and sera from healthy persons, one of the internal clones and five of the C-terminal clones reacted with one or two of the patient sera, but only one clone reacted with all patient sera.
D000907:Antibodies, Bacterial / Q000276:immunology; D000942:Antigens, Bacterial / Q000096:biosynthesis* / Q000235:genetics; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D000939:Epitopes / Q000096:biosynthesis / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D009177:Mycoplasma pneumoniae / Q000235:genetics* / Q000276:immunology; D011019:Pneumonia, Mycoplasma / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D011994:Recombinant Proteins / Q000096:biosynthesis*
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D000939:Epitopes; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins