TABLE 1.
Plasmids used in this study
| Plasmid name | Relevant characteristics | Reference(s) or source |
|---|---|---|
| NAH7 | IncP-9, naphthalene degradation, Tn4655 | 54, 65 |
| pDTG1 | IncP-9, naphthalene degradation, intA gene is nearly identical with tnpI of NAH7 | 10 |
| pSTV29 | Cmr, cloning vector, p15Aori | TAKARA BIO |
| pUC18 | Apr, cloning vector, pMB1ori | 64 |
| pUC19 | Apr, cloning vector, pMB1ori | 64 |
| pUC4K | Apr Kmr, pMB1ori, the Kmr cassette is flanked by EcoRI, BamHI, SalI, and PstI sites | 50 |
| pUC-tnpI | Apr, pUC18 derivative carrying the tnpI gene of Tn4655 | This study |
| pMS0411 | Cmr, pSTV29 derivative carrying the 537-bp fragment located upstream of the tnpI gene (see Fig. 4A and B) at the SalI site | This study |
| pMS0412 | Cmr, pMS0411 derivative carrying another copy of the 537-bp fragment at the KpnI site | This study |
| pMS0413a | Cmr Kmr, pMS0412 derivative in which the Kmr determinant at the BamHI site is flanked by two copies of the 537-bp fragment (see Fig. 4C and 5) | This study |
| pMS0431b | Cmr, pSTV29 derivative carrying the 119-bp attI site between the SacI and KpnI sites (see Fig. 4D) | This study |
| pMS0435 | Cmr, pMS0431 derivative carrying another copy of the 119-bp attI site between the SalI and SphI sites | This study |
| pMS0471 | Cmr Kmr, pMS0435 derivative in which the Kmr determinant at the BamHI site is flanked by two copies of the 119-bp attI site (see Fig. 4C and 5) | This study |
| pMS0477 | Cmr Kmr, pMS0471 derivative in which the 119-bp attI site between the SalI and SphI sites was replaced with a similar fragment from pDTG1 | This study |
| R388c | Tpr Sur, IncW, tra+ | 59 |
| R388attKm | Tpr Kmr, tra+, R388 derivative carrying the 119-bp attI site and a Kmr determinant between the EcoRI and SacI sites | This study |
a
Various deletion derivatives were constructed with PCR to investigate the resolution function of the tnpI gene product and attI site. The names and schematic structures of these derivatives are shown in Fig. 4C.
b
Various deletion derivatives were constructed with PCR to investigate the integration function of the tnpI gene product and attI site. The names and schematic structures of these derivatives are shown in Fig. 4D.
c
Tpr, trimethoprim resistance; Sur, sulfonamide resistance.