Patent ID: 9745572
Date: 2017-08-29
CPC Classifications: A61P,C12N,C40B,G16B

Claim:
1. A method for identifying genes serving as an antibiotic target, including putative essential genes, the method comprising the steps of: (a) generating a pool of mutant bacteria by transposon mutagenesis with an activating transposon (Tn (b) growing bacteria from the mutant pool in the presence of different amounts of an antibiotic to produce two or more test cultures; and (c) comparing the distribution of Tn wherein the method further comprises (d) generating an antibiotic resistant mutant of a bacterium by a method comprising the step of selecting for growth in the presence of an antibiotic to produce an antibiotic resistant mutant clone (Ab R mutant); (e) transforming the Ab R mutant with: (i) one or more wild-type copies of essential genes of said bacterium; and (ii) a transposon which insertionally inactivates bacterial DNA, to produce a pool of transposon mutants which are merodiploid for said one or more essential genes; (f) growing bacteria from the merodiploid pool in the presence of different amounts of said antibiotic to produce two or more test cultures, wherein transposon insertions into an essential gene serving as an antibiotic target in the bacterial DNA are represented under non-selective conditions but not under selective conditions, and wherein one amount of said antibiotic is non-selective and another amount of said antibiotic is selective; and (g) comparing the distribution of transposon insertions between test cultures to identify putative essential genes.