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A Mycobacterium leprae ‐specific gene encoding an immunologically recognized 45 kDa protein
Author(s) -
De Wit Tobias F. Rinke,
ClarkCurtiss Josephine E.,
Abebe Feseha,
Kolk Arend H. J.,
Janson Anneke A. M.,
Agterveld Miranda van,
Thole Jelle E. R.
Publication year - 1993
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.1993.tb00953.x
Subject(s) - biology , mycobacterium leprae , southern blot , microbiology and biotechnology , mycobacterium tuberculosis , polymerase chain reaction , hybridization probe , gene , dna , antibody , genomic dna , lepromatous leprosy , virology , leprosy , tuberculosis , genetics , immunology , medicine , pathology
Summary By screening a Mycobacterium leprae lambda gt11 expression library with a serum from an Ethiopian lepromatous leprosy (LL) patient a clone was isolated (LL4) belonging to hybridization group III of a panel of previously isolated M. leprae clones. Members of this hybridization group encode a serologically recognized 45 kDa protein. The complete DNA sequences of the partially overlapping clones LL4 and L1 (hybridization group III) are presented and these revealed the presence of an open reading frame (ORF) predicting a protein with a molecular size of 42 448 Da. Southern hybridizations on total genomic DNA of M. Ieprae, Mycobacterium tuberculosis and eight atypical mycobacteria showed that the LL4 DNA fragment is specific for M. Ieprae DNA even under low‐stringency conditions. The M. Ieprae specificity of LL4 DNA was further confirmed by the polymerase chain reaction using four different sets of primers. Western blotting analyses showed that the M. Ieprae 45 kDa protein is frequently recognized by antibodies from leprosy patients and that this recognition is specific since no antibodies could be detected in sera of tuberculosis patients. T‐cell proliferation assays also demonstrated T‐cell recognition by leprosy patients and healthy contacts of the M. Ieprae 45 kDa protein. The specificity of the LL4 DNA region and the 45 kDa antigen that is encoded by hybridization group III could provide unique tools for the development of M. Ieprae ‐specific immunological and DNA reagents.