       Document 0557
 DOCN  M9590557
 TI    Phage infection, transfection and transformation of Mycobacterium avium
       complex and Mycobacterium paratuberculosis.
 DT    9509
 AU    Foley-Thomas EM; Whipple DL; Bermudez LE; Barletta RG; Department of
       Veterinary and Biomedical Sciences, University of; Nebraska, Lincoln
       68583-0905, USA.
 SO    Microbiology. 1995 May;141 ( Pt 5):1173-81. Unique Identifier : AIDSLINE
       MED/95291460
 AB    Mycobacterium avium complex strains and Mycobacterium paratuberculosis
       are closely related intracellular pathogens affecting humans and
       animals. M. avium complex infections are a leading cause of morbidity
       and mortality in AIDS patients, and M. paratuberculosis is the agent of
       Johne's disease in ruminants. Genetic manipulation of these
       micro-organisms would facilitate the understanding of their
       pathogenesis, the construction of attenuated vaccine strains and the
       development of new drugs and treatment methods. This paper describes the
       replication of mycobacterial shuttle phasmids and plasmids, and the
       expression of the firefly luciferase reporter gene in M. avium complex
       and M. paratuberculosis. The mycobacteriophage TM4 propagated on M.
       smegmatis or M. paratuberculosis plaqued at the same efficiency on these
       two mycobacterial hosts. Screening of M. avium complex and M.
       paratuberculosis clinical isolates with TM4-derived luciferase reporter
       phages demonstrated that the majority of these isolates were susceptible
       to TM4. Conditions for introduction of DNA were determined by
       transfection of M. paratuberculosis with TM4 DNA and applied to isolate
       kanamycin-resistant transformants of M. avium complex and M.
       paratuberculosis with Escherichia coli-Mycobacterium shuttle plasmids.
       Recombinant plasmids were recovered from transformants without apparent
       loss of DNA sequences. These results provide the basis for the genetic
       manipulation of these pathogenic mycobacterial species.
 DE    Amikacin/PHARMACOLOGY  Animal  AIDS-Related Opportunistic
       Infections/VIROLOGY  Bacteriophages/*GENETICS  Comparative Study  Human
       Kanamycin/PHARMACOLOGY  Kinetics  Luciferase/ANALYSIS/BIOSYNTHESIS
       Microbial Sensitivity Tests  Mycobacterium avium Complex/DRUG
       EFFECTS/*GENETICS/*VIROLOGY  Mycobacterium paratuberculosis/DRUG
       EFFECTS/*GENETICS/*VIROLOGY  Plasmids  Recombinant
       Proteins/ANALYSIS/BIOSYNTHESIS  Species Specificity  Support, Non-U.S.
       Gov't  Support, U.S. Gov't, Non-P.H.S.  Support, U.S. Gov't, P.H.S.
       Transfection  *Transformation, Bacterial  Tuberculosis/ETIOLOGY/VIROLOGY
       JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).

