Cloning and expression in Escherichia coli of Mycoplasma gallisepticum antigens recognized by sera from infected chickens.
A clone bank of Mycoplasma gallisepticum (MG) strain A5969 DNA was prepared in the expression vector phage lambda gt11. Approximately 75% of the resulting phages were recombinants, based upon the insertional inactivation of the lacZ gene of the vector. Clones were screened immunologically with serum prepared from specific-pathogen-free white leghorn chickens that had been infected with aerosolized MG. Approximately 250 clones, or less than 1% of the recombinant phage, reacted positively to various degrees with the test serum and failed to react with serum from uninfected specific-pathogen-free control chickens. A single clone was chosen at random for comparison with a vector control by western immunoblot, revealing a polypeptide of 140,000 molecular weight in the clone profile but not the control profile that reacted with immune serum. Clones expressing MG antigens recognized during infection may provide an improved means for antigen preparation for serologic diagnosis of mycoplasmosis.
Krause
D C
DC
Department of Microbiology, University of Georgia, Athens 30602.
Kleven
S H
SH
Lee
K K
KK
D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics* / Q000276:immunology; D010582:Bacteriophage lambda; D015153:Blotting, Western; D002645:Chickens; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D005822:Genetic Vectors; D007106:Immune Sera / Q000276:immunology; D009174:Mycoplasma / Q000235:genetics / Q000276:immunology*; D009175:Mycoplasma Infections / Q000276:immunology / Q000662:veterinary; D011201:Poultry Diseases / Q000276:immunology; D013047:Specific Pathogen-Free Organisms
D000942:Antigens, Bacterial; D007106:Immune Sera