ABSTRACT
A recombinant phage library harbouring Mycoplasma meleagridis (MM) genomic DNA fragments was generated in the bacteriophage lambda gt11 expression vector. The library was screened for expression of MM specific antigens with a polyclonal antiserum that had been preadsorbed with antigens of the most common unrelated avian mycoplasma species. A 49-amino acid antigenic domain unique to MM was isolated, expressed in Escherichia coli, and its serodiagnostic potential was demonstrated. An antiserum raised against this MM-specific antigenic domain recognized a cluster of seven membrane-associated MM proteins with molecular masses ranging from 34 to 75 kDa. Overall, this study resulted in the identification of a potent serodiagnostic tool and revealed the complex antigenic nature of MM.
Subject(s)
Antigens, Bacterial/genetics , Cloning, Molecular , Mycoplasma meleagridis/genetics , Mycoplasma meleagridis/immunology , Animals , Antibodies, Bacterial/metabolism , Antigens, Bacterial/biosynthesis , Antigens, Bacterial/immunology , Base Sequence , DNA, Bacterial/chemistry , Escherichia coli/genetics , Immune Sera/metabolism , Immunoblotting/methods , Molecular Sequence Data , Mycoplasma Infections/diagnosis , Mycoplasma Infections/veterinary , Poultry Diseases/diagnosis , Poultry Diseases/microbiology , Species Specificity , TurkeysABSTRACT
To determine whether insulin stimulates human ovarian testosterone production in the polycystic ovary syndrome by activating its own receptor and using inositolglycan mediators as the signal transduction system, thecal cells from polycystic ovary syndrome women were isolated and cultured. Insulin and insulin-like growth factor I stimulated thecal testosterone biosynthesis. Antibody blockade of the insulin receptor abolished insulin's stimulatory action, whereas effective antibody blockade of the insulin-like growth factor I receptor did not alter insulin's stimulation of thecal testosterone biosynthesis. A chiro-inositol containing glycan (INS-2) increased thecal testosterone biosynthesis. Preincubation of cells with an antiinositolglycan antibody (A23939 or alpha IGP) abolished insulin's stimulatory effect, but not that of hCG. These findings suggest that inositolglycans serve as the signal transduction system for insulin's stimulation of human thecal testosterone biosynthesis.
