TABLE 1.
Bacterial strains, plasmids, and oligonucleotide primers used in this study
| Strain, plasmid, or primer | Source and/or reference; description or sequence (5′-); restriction sitea |
|---|---|
| Bacterial strains | |
| Acinetobacter sp. strain DSM 17874 | 24; Camr, deposited in the DSMZ strain collection under accession no. DSM17874 |
| Escherichia coli S17-1 (λpir) | 4 |
| Acinetobacter baylyi ADP1 | A. Steinbüchel, 8 |
| Acinetobacter sp. strain M-1 | Y. Sakai, 21 |
| Acinetobacter sp. strain RAG-1 | Purchased from ATCC, accession no. ATCC31012 |
| Acinetobacter sp. strain MAV1 | This study; almA knockout mutant, Camr Aprr |
| Acinetobacter sp. strain BKO2 | This study; almB knockout mutant, Camr Aprr |
| Plasmids | |
| pLOFKm | 7; mini-Tn10 delivery vector, Kanr |
| pSOK201 | 28; Aprr |
| pSOK804 | 3; Aprr |
| pDLM02.1 | 13; A. baylyi ADP1 and ColE1 replication origin, Tdk/Kanr selection/counter-selection cassette, Ampr |
| pLAL50 | This study; part of pSOK804 containing a 607-bp almA fragment, Aprr |
| pBKO2 | This study; derivative of pSOK201 containing a 300-bp orf1 fragment, Aprr |
| pALMA1 | This study; derivative of pDLM02.1 carrying the almA gene from Acinetobacter sp. strain 6A2, Ampr Kanr |
| pACIAD1 | This study; derivative of pDLM02.1 carrying the ACIAD3192 gene from A. baylyi ADP1, Ampr Kanr |
| Primers | |
| Tn10:1 | GGATCATATGACAAGATGTG |
| Kan1 | GCTCTAGACCGTCAAGTCAGCGTAATGC |
| almA1 | GACATGTGTATTGTCAAATTTGTGC |
| almA2 | CCAATGAGATCATGGAAGAAC |
| almA3 | GCTCTAGACTATCCTGGTATTCGTTCAG; XbaI |
| almA4 | CGGGATCCTAAATACCACGTTGCATACC; BamHI |
| almA5 | GCGCGCGCATGCGACATGTGTATTGTCAAATTTGTGC; SphI |
| almA6 | GCGCGCGGCGCCCCCTATGCCATGCATAGGGTTTC; NarI |
| orf1:1 | GCGCGCGCATGCGACAGCAATTGCTGAGAAGTTGG; SphI |
| orf1:2 | GCGCGCGAATTCGTGAAATGAGCAACCTCTCCTCC; EcoRI |
| ACIAD1 | GCGCGCGCATGCCGCGAAGTAAACACTTCAGCAG; SphI |
| ACIAD2 | GCGCGCGGCGCCCCTTTTTTTGTTTAGCTCAAGTTAGGATAC; NarI |
a
Antibiotic resistance markers: Ampr, ampicillin resistance; Camr, chloramphenicol resistance; Kanr, kanamycin resistance; Aprr, apramycin resistance. Restriction sites are underlined.