TABLE 1.
Strains, plasmids, and primers used in this study
| Strain, plasmid, or primer | Relevant characteristic(s) or sequence (5′ to 3′) | Reference or usage |
|---|---|---|
| G. sulfurreducens | ||
| Wild type | ATCC 51573 | 12 |
| 20B5 | pMiniHimar RB1 insertion at 1646730, Kmr | 61 |
| 67B2 | pMiniHimar RB1 insertion at 1646910, Kmr | 61 |
| Δ1501 | Kanamycin resistance gene replacing GSU1501, Kmr | This study |
| E. coli | ||
| WM3064 | Donor strain for conjugation: thrB1004 pro thi rpsL hsdS lacZΔM15 RP4-1360 Δ(araBAD)567 ΔdapA1341::[erm pir(wt)] | 64 |
| Plasmids | ||
| pBBR1MCS-2 | Mobilizable broad-host-range plasmid; lacZ, Kmr | 31 |
| pBBR1MCS-5 | Mobilizable broad-host-range plasmid; lacZ, Gmr | 31 |
| pGCOMP1500-5 | GSU1500 to GSU1505 in MCS of pBBR1MCS-5, Gmr | This study |
| Primers | ||
| JR03F | TGATCTGTTGGAATACGGGATGAAGTACCG | PCR amplification of sequence upstream of GSU1501 |
| JR04R | GAAATTCCCCTCTTCAGTTATTCTGAACTC | |
| JR05F | TTGTGAGCTTCCAAGCATTTCTAAGGATCT | PCR amplification of sequence downstream of GSU1501 |
| JR06R | CACACTGAGGAAGAGCCAGCTCGATATCGG | |
| JR07F | TAACTGAAGAGGGGAATTTCAGCGAACCGGAATTGCCAGCT | PCR amplification of Kanr from pBBR1MCS-2, with overhangs homologous to JR04R and JR05F in italics |
| JR08R | GAAATGCTTGGAAGCTCACAATCAGAAGAACTCGTCAAGAAGGC | |
| JR09F | GTCCAACAAAGGGAACTGCT | qPCR detection of GSU1501 |
| JR10R | CCTCCGCAGAGAGGTAATCA | |
| JR11F | AGTTCTCGACGTACGCCACT | qPCR detection of rpoD |
| JR12R | TCAGCTTGTTGATGGTCTCG | |
| JR13F | CACCGGCATAATCTCCAAGT | qPCR detection of recA |
| JR14R | TTGAGGGATGCGATCTTGCG | |
| JR15F | TACTCTAGAAATGGTGCCGTTCACCGACCA | PCR amplification of GSU1500 to GSU1505; XbaI and SacI restriction sites in italics |
| JR16R | GACGAGCTCTCATAAACGAACCTCGTCCC |