D29 shuttle phasmids and uses thereof

Details D29 shuttle phasmids and uses thereof D29 shuttle phasmids and uses thereof

1996-03-07

1998-06-30

Degen, Nancy

Chemistry: molecular biology and microbiology

Treatment of micro-organisms or enzymes with electrical or...

Modification of viruses

4352523, 4353201, 536 231, C12N 121, C12N 1511, C12N 1563, C12N 1574

Patent

active

057732671

ABSTRACT:
The present invention provides a conditional shuttle phasmid constructed by inserting a cosmid into a non-essential region of the D29 mycobacteriophage which is capable of introducing DNA of interest into the chromosome of mycobacteria, especially M. tuberculosis complex organisms and other slow growing mycobacteria. The present invention provides a recombinant mycobacterium which expresses a DNA of interest incorporated into its chromosome by a conditional shuttle plasmid containing the DNA of interest. The present invention further provides a mycobacterial auxotrophic mutant and method of generating auxotrophic mutants. Finally, the present invention provides a method of inactivating a mycobacterial virulence gene.

REFERENCES:
Sellers et al. (1970) The effects of ultraviolet irradiation on mycobacteriophages and their infectious DNAs. J. Gen. Virol. 7:233-247, Jun. 1970.
Jacobs et al. (1989) Mycobacteriophage vector systems. Rev. Infect. Dis. 11, Supp. 2:S404-S410, Mar. 1989.
Sambrook et al. (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, 13.1-13.102, 1989.

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