Detection and characterization of a lambda gt11 recombinant clone of M. leprae that expresses an antigenic determinant of a 64-kDa protein.
A genomic library of Mycobacterium leprae in the expression vector lambda gt11 was screened with rabbit polyclonal hyperimmune antiserum elicited with a sonicated extract of M. leprae. Numerous reactive clones were isolated by this immunoscreening, indicating a broad antibody response of the rabbit. None of the recombinant clones was reactive with monoclonal antibodies to the previously well-characterized M. leprae recombinant clones. One of the clones isolated, clone A, encoded a large, 132-143-kDa beta-galactosidase fusion protein expressing an epitope of M. leprae. Monospecific antibodies eluted from this fusion protein reacted on Western blots with a 64-kDa M. leprae protein. The DNA of this clone was shown to be distinct from the gene encoding the well-characterized immunodominant 65-kDa protein by DNA hybridization. We have identified a new lambda gt11 recombinant clone encoding a fusion protein with a 64-kDa protein. This protein is recognized by the humoral immune response of the rabbit in response to a challenge with an M. leprae cell extract.
Davidson
S K
SK
Department of Microbiology and Immunology, Morehouse School of Medicine, Atlanta, Georgia 30310.
George
G T
GT
D000942:Antigens, Bacterial / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D001435:Bacteriophages; D015139:Blotting, Southern; D015153:Blotting, Western; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000032:analysis / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000032:analysis / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D015723:Gene Library; D006801:Humans; D008242:Lysogeny; D009166:Mycobacterium leprae / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics
D000942:Antigens, Bacterial; D004269:DNA, Bacterial; D000939:Epitopes; D011993:Recombinant Fusion Proteins