Reporter gene based method for the screening of anti-tuberculosis drugs by using essential and regulatory genes of mycobacteria as drug target
AssigneeCouncil of Scientific and Industrial Research
InventorsVishal Soni, Lakshami Pathi Khandrika, Pushpa Agrawal
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Abstract
The present invention relates to a method for making recombinant Saccharomyces cerevisiae. The method includes the steps of amplifying one or more whiB-like genes of Mycobacteria by polymerase chain reaction (PCR), cloning the amplified one or more whiB-like genes into a plasmid, transforming the clone into an E. coli using a first shuttle vector, amplifying the clone, introducing the amplified clone into a second shuttle vector, introducing said second shuttle vector into Saccharomyces cerevisiae.
Claims16
A method for making a recombinant Saccharomyces cerevisiae said method comprising the steps of:
- a) amplifying one or more whiB-like genes of Mycobacteria by polymerase chain reaction (PCR), wherein the whiB-like genes are selected from the group consisting of whiB1 (SEQ ID NO:1), whiB2 (SEQ ID NO:3), whiB3 (SEQ ID NO:5), and whiB4 (SEQ ID NO:7) and the Mycobacteria are selected from the group consisting of Mycobacterium tuberculosis, Mycobacterium bovis, Mycobacterium avium, Mycobacterium fortuitum, Mycobacterium gastri, Mycobacterium kansasii, Mycobacterium leprae, Mycobacterium marinum, Mycobacterium microti, Mycobacterium phlei, Mycobacterium scrofulaceum, Mycobacterium smegmatis, and Mycobacterium xenopi,
- b) cloning the amplified one or more whiB-like genes into a plasmid,
- c) transforming the clone into an E. coli using a first shuttle vector,
- d) amplifying the clone,
- e) introducing the amplified clone into a second shuttle vector,
- f) introducing the second shuttle vector into Saccharomyces cerevisiae, and
- g) obtaining transformed Saccharomyces cerevisiae.
The method as claimed in claim wherein the DNA-binding proteins repress the activation of β-galactosidase.
Dependencies are inferred from the claim text.