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Published on: October 14, 2011
Isolation and characterization of recombinant lambda gt11 bacteriophages expressing four different Mycobacterium
S L Morris1, D A Rouse, D Hussong
1Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland 20892.
Abstract:
Four bacteriophages expressing different immunoreactive recombinant Mycobacterium intracellulare antigens were isolated from a lambda gt11 library with monoclonal antibodies to M. intracellulare. These four antibodies reacted with native M. intracellulare 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. intracellulare derivation of the inserts. The physical maps of the immunoreactive phages were deduced by restriction enzyme digestions. The molecular weights of the expressed recombinant antigens were determined by Western (immuno-) blotting.
Insights
Researchers identified four bacteriophages containing Mycobacterium intracellulare antigens. These phages express key immunoreactive proteins, aiding in the study of M. intracellulare and potential diagnostic development.
Area of Science:
- Microbiology
- Immunology
- Molecular Biology
Background:
- Mycobacterium intracellulare is an important human pathogen.
- Identifying specific antigens is crucial for diagnostic and therapeutic development.
Purpose of the Study:
- To isolate and characterize recombinant Mycobacterium intracellulare antigens expressed by bacteriophages.
- To confirm the origin and analyze the properties of these immunoreactive antigens.
Main Methods:
- Screening a lambda gt11 library using monoclonal antibodies against M. intracellulare.
- Southern blot hybridizations to confirm DNA insert origin.
- Restriction enzyme digestion for physical mapping of phages.
- Western (immuno-) blotting to determine molecular weights of expressed antigens.
Main Results:
- Four bacteriophages expressing immunoreactive M. intracellulare antigens were isolated.
- The identified antigens correspond to native M. intracellulare proteins of 54, 43, 40/38, and 22 kDa.
- DNA inserts were confirmed to be of M. intracellulare origin.
- Physical maps and molecular weights of recombinant antigens were determined.
Conclusions:
- Successfully isolated and characterized novel recombinant antigens from M. intracellulare.
- These findings provide valuable tools for further research into M. intracellulare infections.
- The identified antigens hold potential for developing diagnostic assays.
