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Use Of The Caulobacter Crescentus Genome Sequence To Develop A Method For Systematic Genetic Mapping, Craig Stephens, Lisandra West, Desiree Yang
Use Of The Caulobacter Crescentus Genome Sequence To Develop A Method For Systematic Genetic Mapping, Craig Stephens, Lisandra West, Desiree Yang
Biology
The functional analysis of sequenced genomes will be facilitated by the development of tools for the rapid mapping of mutations. We have developed a systematic approach to genetic mapping in Caulobacter crescentus that is based on bacteriophage-mediated transduction of strategically placed antibiotic resistance markers. The genomic DNA sequence was used to identify sites distributed evenly around the chromosome at which plasmids could be nondisruptively integrated. DNA fragments from these sites were amplified by PCR and cloned into a kanamycin-resistant (Kanr) suicide vector. Delivery of these plasmids into C. crescentus resulted in integration via homologous recombination. A set of 41 strains …