Abstract
Mycobacteria are major pathogens of man and animals. There are ˜10 million cases of tuberculosis world wide with an annual mortality of three million1 people. Leprosy, caused by Mycobacterium leprae, afflicts over ten million people, primarily in developing countries2. M. tuberculosis and mycobacteria of the M. aviumintracellulare-scrofulaceum (MAIS) group are major opportunistic pathogens of patients with acquired immune deficiency syndrome (AIDS)3. M. paratuberculosls is the cause of Jöhne's disease in cattle. Yet, BCG (Bacille Calmette-Guerin), an avirulent strain of M. bovis, is the most widely used human vaccine in the world, having been administered to about 2.5 × 109 people since 1948 (ref. 4). BCG was highly protective against tuberculosis in England5, but has been found not to be effective in preventing pulmonary tuberculosis in adults in Southern India6. We have initiated studies to develop the methodology for efficient gene transfer in mycobacteria. We have constructed recombinant shuttle phasmids which are chimaeras containing mycobacteriophage DNA into which an E. coli cosmid is inserted. They can replicate in E. coli as plasmids and in mycobacteria as phages, and transfer DNA across both genera. These shuttle vectors permit for the first time the introduction of foreign DNA by infection into M. smegmatis and BCG. By introducing and ultimately expressing genes for protective antigens for a variety of pathogens, it may be possible to develop cultivatable mycobacteria into useful multivaccine vehicles.
References
1. Joint International Union Against Tuberculosis and World Health Organization Study Group Tubercle 63, 157–169 (1982). 2. Bloom, B. R. & Godal, T. Rev. infect. Dis. 5, 765–780 (1983). 3. Centers for Disease Control Morbid. Mortal, wkly. Rep. 34, 774 (1986). 4. United Nations Children's Emergency Fund (UNICEF) The State of the World's Children 131 (Oxford University Press, 1985). 5. Hart, P. D. & Sutherland, I. Br. Med. J. 2, 293–295 (1977). 6. Tuberculosis Prevention Trial, Madras Ind. J. med. Res. 72 (suppl.), 1–74 (1980). 7. Okanishi, M., Suzuki, K. & Umezawa, H. /. gen. Microbiol. 80, 389–400 (1974). 8. Hopwood, D. A. & Wright, H. M. Molec. Gen. Genet. 162, 307–317 (1978). 9. Bibb, M. J., Ward, J. M. & Hopwood, D. A. Nature 274, 398–400 (1978). 10. Crawford, J. T. & Bates, J. H. Infect. Immunity 24, 979–981 (1979). 11. Mizuguchi, Y., Fukunaga, M. & Taniguchi, H. /. Bact. 146, 656–659 (1981). 12. Meissner, P. S. & Falkinham, J. O. Ill J. Bact. 157, 669–672 (1984). 13. Labidi, A., David, H. L. & Roulland–Dussoix, D. Annls Microbiologie 136B, 209–215 (1985). 14. Brenner, S., Cesareni, G. & Karn, J. Gene 17, 27–44 (1982). 15. Suarez, J. E. & Chater, K. F. Nature 286, 527–529 (1980). 16. Timme, T. L. & Brennan, P. J. /. gen. Microbiol. 130, 2059–2066 (1984). 17. Collins, J. & Hohn, B. Proc. natn. Acad. Sci. U.S.A. 75, 4242–4246 (1978). 18. Hohn, B. & Collins, J. Gene 11, 291–298 (1980). 19. Lotte, A. et al. Adv. Tuberc. Res. 32, 107–193 (1984). 20. Hopwood, D. A. et al. In Genetic Manipulation of Streptomyces–A Laboratory Manual (John Innes Foundation, Norwich, 1985). 21. Froman, S., Will, D. W. & Bogen, E. Am. J. Public Health 44, 1326–1334 (1954). 22. Redmond, W. B. & Cater, J. C. Am. Rev. resp. Dis. 82, 781–787 (1960). 23. Udou, T., Ogawa, M. & Mizuguchi, Y. /. Bact. 151, 1035–1039 (1982). 24. Birnboim, H. C. & Doly, J. Nucleic Acids Res. 7, 1513–1525 (1979). 25. Benton, W. D. & Davis, R. W. Science 196, 180–182 (1977). 26. Southern, E. / molec. Biol. 98, 503–517 (1975). 27. Jones, W. D. Jr Tubercle, Lond. 60, 55–58 (1979). 28. Jones, W. D. Jr & Greenberg, J. /. din. Microbiol. 7, 467–469 (1978).
Author information
Affiliations
Rights and permissions
About this article
Cite this article
Jacobs, W., Tuckman, M. & Bloom, B. Introduction of foreign DNA into mycobacteria using a shuttle phasmid. Nature 327, 532–535 (1987). https://doi.org/10.1038/327532a0
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/327532a0
Further reading
-
Engineered Bacteriophage Therapeutics: Rationale, Challenges and Future
BioDrugs (2021)
-
Genome-wide identification of essential genes in Mycobacterium intracellulare by transposon sequencing — Implication for metabolic remodeling
Scientific Reports (2020)
-
Mycobacterium Lysine ε-aminotransferase is a novel alarmone metabolism related persister gene via dysregulating the intracellular amino acid level
Scientific Reports (2016)
-
Recombinant Mycobacterium bovis BCG for immunotherapy in nonmuscle invasive bladder cancer
Applied Microbiology and Biotechnology (2015)
-
Development of rapid microwave-mediated and low-temperature bacterial transformations
Journal of Chemical Biology (2013)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.
