TABLE 1.
Biological materials used in this worka
| Strain or plasmid | Relevant characteristic(s) | Strain construction or reference |
|---|---|---|
| Strains | ||
| E. coli | ||
| DH5α | recA1 ΔlacZ | Invitrogen |
| S17-1 | Smr Tprmod+res thi pro recA hsdR17, integrated plasmid RP4-TC::Mu-Km::Tn7 into genome | 28 |
| AKN67 | pBK-mini-Tn7gfp3 Kmr | 19 |
| AKN132 | pBK-mini-Tn7dsred Gmr Cmr | 33 |
| AKN133 | Mini-Tn7Km-dsred Kmr Smr Cmr | 33 |
| C. crescentus | ||
| CB15 | Wild type; Ampr | 24 |
| NA1000 | syn-1000, CB15 synchronizable derivative, unable to form holdfast and rosettes | 11 |
| LS801 | flgF::Tn5 Kmr in CB15 | L. Shapiro, unpublished data |
| LS1088 | CC0095::Tn5, Kmr in CB15, holdfast deleted | L. Shapiro, unpublished data |
| PV14 | ΔpilA-cpaF::Ωaac3, Aprr in NA1000 | 35 |
| PV35 | ΔpleA::Δaac3, Aprr in NA1000 | 34 |
| PV1735 | ΔpodJ in CB15 | 35 |
| SC268 | motA102 point mutation in CB15 | 38 |
| SC286 | motB108 point mutation in CB15 | 38 |
| SC1035 | pleC::Tn5 Kmr in CB15 | 36 |
| SC1119 | podJ::Tn5 Kmr in CB15 | 36 |
| YB2878 | hfsA125 Kmr in CB15 | 31 |
| AS100 | flgH::Tn5 Kmr in CB15 | This work |
| AS109 | miniTn7Km-dsred, Kmr in CB15 | This work |
| AS110 | pBK-mini-Tn7gfp3 Kmr in CB15 | This work |
| AS111 | ΔpilA-cpaF::Ωaac3 gfp3 Kmr Aprr | φCr30 (PV14) × AS110, selected on Apr |
| AS112 | ΔpilA-cpaF::Ωaac3 motA gfp3 Kmr, Aprr | φCr30 (PV14) × AS115, selected on Apr |
| AS113 | pBK-mini-Tn7dsred Kmr Gmr in YB2878 | This work |
| AS114 | pBK-mini-Tn7dsred Kmr Gmr in AS100 | This work |
| AS115 | pBK-mini-Tn7gfp3 Kmr in SC268 | This work |
| AS116 | flgH::Tn5 ΔpilA-cpaF::Ωaac3 gfp3 Kmr Aprr Testr | φCr30 (AS100) × AS111, selected on Tet |
| AS117 | ΔpilA-cpaF::Ωaac3 hfsA dsred Kmr Gmr Aprr | φCr30 (PV14) × AS113, selected on Apr |
| AS118 | flgH::Tn5 hfsA dsred Kmr Gmr Tetr | φCr30 (AS100) × AS113, selected on Tet |
| AS119 | motA flgH::Tn5 Kmr | φCr30 (AS100) × SC268, selected on Km |
| AS120 | In-frame deletion of CC0095 in CB15 | This work |
| AS121 | pBK-mini-Tn7gfp3 Kmr in AS120 | This work |
| Plasmids | ||
| pBluescript KS | AprlacPOZ | Stratagene |
| pNPTS138 | pLitmus38-derived vector with oriT, sacB, and nptI genes, integrates into Caulobacter genome, Kmr | M. R. K. Alley, unpublished data |
| pNPT228 | pLitmus28-derived vector with oriT and nptII genes, Kmr | M. R. K. Alley, unpublished data |
| pHP45Ωaac3 | Source of omega Aprr cassette | 5 |
| pCRtopo4 | Topoisomerase I bound covalently to the 3′-T overhangs, Kmr Ampr | Invitrogene |
| pBK-mini-Tn7gfp3 | Delivery plasmid for miniTn7-gfp3; mob+ Kmr Cmr Smr | 19 |
| pBK-miniTn7dsred | Delivery plasmid for miniTn7-dsred; mob+ Gmr Cmr | 33 |
| miniTn7Km-dsred | Delivery plasmid for miniTnT-dsred; mob+ Kmr Smr Cmr | 33 |
| pRK600 | Cmrori-ColE1 RK2-mob+ RK2-tra+ helper plasmid | 17 |
| pUX-BF13 | Amprmob+ori-R6K; helper plasmid | 3 |
| pPE305 | pCRtopo4::3′ end of CC0094 into 5′ end of CC0095 | This work |
| pPE306 | pCRtopo4::3′ end of CC0095 into 5′ end of CC0096 | This work |
| pPE307 | pNTPS138 with EcoRI-KpnI fragment containing deletion in CC0095 | This work |
a
Amp,r, ampicillin resistance; Kmr, kanamycin resistance; Gmr, gentamycin resistance; Tetr, tetracycline resistance; Aprr, apramycin resistance; Cmr, chloramphenicol resistance; Smr, streptomycin resistance.