Isolation and characterization of recombinant lambda gt11 bacteriophages expressing four different Mycobacterium intracellulare antigens

Abstract

Four bacteriophages expressing different immunoreactive recombinant Mycobacterium intracellulare antigens were isolated from a Agtll library with monoclonal antibodies to M. intracellulare. These four antibodies reacted with native M. intracelulare proteins of 54, 43, 40/38, and 22 kilodaltons. Southern blot hybridizations with DNA probes prepared from insert fragments of these bacteriophages confirmed the M. intracelulare derivation of the inserts. The physical maps of the inmunoreactive phages were deduced by restriction enzyme digestions. The molecular weights of the expressed recombinant antigens were determined by Western (immuno-) blotting.

Mycobacterium avium complex disease in patients with AIDS: seroreactivity to native and recombinant mycobacterial antigens

“…All of the recombinant proteins were identified from studies of mycobacterial Xgtll gene expression libraries. Five of the recombinant antigens were derived from M. intracellulare antigens of 22, 40, 43, 54, and 85 kDa; three of the proteins were from M. avium antigens of 26, 33, and 34 kDa; and the remaining recombinant protein was from an M. kansasii antigen of 35 kDa (19,20,24). The genes encoding six of these antigens (22, 34, 35, 40, 43, and 85 kDa) were cloned into the pGEMEX expression system (Promega, Madison, Wis.).…”

Duplication of genes encoding the immunodominant 38 kDa antigen in Mycobacterium intracellulare

“…Subsequently, the M. tuberculosis 38 kDa gene probe was found to hybridise to a single PvuII DNA fragment from several isolates classified by molecular typing as M. intracellulare, but not to DNA from several M. avium isolates [3]. To date, only a few genes encoding major antigens of M. intracellulare have been characterised, some partially [6] and others entirely [7-91. In view of the possible role of the 38 kDa antigen of M. intracellulare in some mechanisms of pathogenesis which are not shared with M. avium, we decided to clone and sequence the gene encoding this antigen. We considered that its sequencing will also aid the identification of species-specific epitopes, as a potential aid for the immunodiagnosis of A4.…”

Production, characterization, and species specificity of monoclonal antibodies to Mycobacterium avium complex protein antigens

“…Studies of recombinant M. tuberculosis and Mycobacterium leprae proteins and their genes have greatly expanded our understanding of the immunogenicity of the tubercle and leprae bacilli (7,13,19,22). We have recently described the isolation of four Xgtll clones which express recombinant M. intracellulare proteins by using anti-M. intracellulare MAbs (16). We are confident that the anti-M. avium MAbs characterized in this report will also be useful for screening mycobacterial expression libraries.…”

“…We have recently prepared several MAbs which bind to proteinassociated epitopes of the MAC. The characterization of four M. intracellulare-reactive MAbs, two of which apparently react with MAC-specific epitopes on 40-and 43-kDa proteins, has been described previously (16). In this report, we describe the production and characterization of eight MAbs to M. avium determinants.…”