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Molecular genetics is one of the most powerful tools available to microbiologists to investigate the basis of bacterial phcnotypcs. Even where biochcmical or structural data arc scarcc, genetic analysis (particularly whcn combincd with DNA sequencing) often reveals considerable insight into thc mechanisms involved in microbial processcs of medical, industrial, and cnvironrncntal interest. Current genetic assets avallablc for studying the properties of different microorganisms have been reviewed in Volume 204 of this series and also this volume and elsewhere. 1, 2 In this chaptcr, we discuss the use of minitransposons, developed in our laboratory, for the analysis, construction, and manipulation of complcx phcnotypcs in a widc range of gram-negative bacteria. 3-5 The only two conditions limiting the usc of these minitransposons is that a target strain should (I) bc an cffcctivc recipient in RP4-mcdiatcd conjugal transfer …