Insertional mutations in mycobacteria
AssigneeAlbert Einstein College of Medicine of Yeshiva University
InventorsWilliam R. Jacobs JR., Barry Bloom, Ganjam V. Kalpana, Jeffrey D. Cirillo, Ruth McAdam
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Abstract
A mutated mycobacterium selected from the class consisting of mutated M. bovis-BCG, mutated M. tuberculosis, and mutated M. leprae. The mutation of M. bovis-BCG, M. tubeiculosis, or M. leprae is preferably effected through an insertional mutation of a mycobacterial gene. The insertional mutagenesis may be effected, for example, through illegitimate recombination or by a mycobacterial transposon. Such mutated mycobacteria may then be transformed with an expression vector(s) containing a complement gene to the gene which is mutated, and preferably also including a heterologous gene.
Claims30
A mutated mycobacterium selected from the class consisting of mutated M. bovis-BCG, mutated M. tuberculosis, and mutated M. leprae.
A transformed mutated mycobacterium selected from the class consisting of mutated M. bovis-BCG and mutated M. tuberculosis, said mutated mycobacterium capable of expressing a heterologous gene.
Mutated M. bovis-BCG deposited as ATCC No. 55202.
Mutated M. bovis-BCG deposited as ATCC No. 55203.
A mutated M. bovis wherein said mutation is effected through an insertional mutation of an M. bovis gene.
A transformed mutated mycobacterium selected from the class consisting of mutated M. bovis-BCG, mutated M. tuberculosis, and mutated M. leprae, wherein said mycobacterium is mutated through an insertional mutation of a mycobacterial gene, and wherein said mycobacterium is transformed with DNA including a non-mutated gene which complements said insertionally mutated mycobacterial gene, whereby said mutated mycobacterium is capable of expressing said complementing gene.
The mycobacterial transposon IS1096 as shown in FIG. 29.
A mutated mycobacterium wherein said mycobacterium is mutated by mycobacterial transposon IS1096 as shown in FIG. 29, which inserts randomly into a mycobacterial chromosome to effect an insertional mutation of a mycobacterial gene.
A vector for inserting DNA into the chromosome of a mycobacterium, said vector including a pair of inverted repeat sequences and DNA encoding a transposase, wherein said inverted repeat sequences and said DNA encoding a transposase are the inverted repeat sequences and the transposase sequence of IS1096 as shown in FIG. 29.
A method of mutating BCG, comprising:
- transfecting into a mycobacterium a vector including a pair of inverted repeat sequences and DNA encoding a transposase.
A method of mutating mycobacteria, comprising:
- transfecting into a mycobacterium a vector including a pair of inverted repeat sequences and DNA encoding a transposase, said inverted repeat sequences and said DNA encoding a transposase being the inverted repeat sequences and the transposase sequence of IS1096 as shown in FIG. 29.
Dependencies are inferred from the claim text.