FIG. 4.
pMT266 derivatives carrying tnpA and tnpR genes of Tn4656. The abbreviations for restriction sites are the same as those described in the legends to Fig. 2 and 3. (A) Construction of the pMT2827 derivatives. The derivatives are not drawn to scale. The thin line indicates the pMT266 portion. The open and shaded boxes represent the pRME1-derived fragment and the fragment deleted in the descendant plasmid, respectively. (B) Localization of tnpA and tnpR on pMT3020. Only the Tn4656 portion is shown. Plasmids pMT3031, pMT3032, pMT3034, pMT3035, and pMT3036 are derived from pMT3030, and plasmids pMT3042, pMT3043, and pMT3045 are derived from pMT3041. The thin lines and solid triangles in the pMT3020 derivatives represent the deleted fragment and the insert of the pUC4K-derived Kmr gene, respectively. Note that deletions in pMT3032, pMT3041, and their derivatives extend to the unique NruI site in the pMT266 portion. pMT3020 and its derivatives were examined to determine their ability to complement the defect in cointegration of Tn4656-2939 and the defects in resolution of the Tn4656-2944- and Tn4656-2948-mediated cointegrate (Fig. 3 and 5). For an explanation of the plus and minus signs see the legend to Fig. 3. NT, not tested.