METHOD FOR IDENTIFYING ANTIBIOTIC TARGETS BY COMPLEMENTED SEQUENCING

摘要

Disclosed is a method for identifying an essential gene which serves as an antibiotic target in a bacterium, the method comprising the steps of: (a) generating an antibiotic resistant mutant of said bacterium by a method comprising the step of selecting for growth in the presence of said antibiotic to produce an antibiotic resistant mutant clone (AbR mutant); (b) transforming the AbR mutant with: (i) one or more 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; (c) growing bacteria from the merodiploid pool in the presence of different amounts of said antibiotic to produce two or more test cultures; and (d) comparing the distribution of transposon insertions between test cultures to identify a putative essential gene serving as a target of said antibiotic in said bacterium.