| 摘要 |
A method for the identification of a conditional essential gene of an organism, which method comprises: (i) providing a library of transposon mutants of the said organism (input library); (ii) subjecting a sample of the input library to a conditional restraint in order to generate an output library; (iii) isolating oligonucleotide sequences flanking transposon insertion sites from the input and output libraries to give input and output pools of oligonucleotide sequences; (iv) hybridising the input and output pools of oligonucleotide sequences with the same oligonucleotide microarray or separately with identical olionucleotide microarrays, wherein the oligonucleotide microarray comprises oligonucleotide probes from the organism of step (i) and/or oligonucleotide probes having at least 80% sequence identity thereto; and (v) calculating an attenuation ratio for an oligonucleotide probe on the oligonucleotide microarray by determining the ratio of the signal from hybridisation of the output pool of flanking sequences to said oligonucleotide probe to the signal from hybridisation of the input pool of flanking sequences to said oligonucleotide probe, wherein an attenuation ratio of less than I is indicative of an oligonucleotide probe on the oligonucleotide microarray which corresponds to a sequence derived from a transposon insertion into a conditional essential gene. Genes identified in such a method may be useful in the preparation of a vaccine and may themselves be targets for drug discovery. |
