TABLE 2.

Primers used in the plasmid backbone analysisa

Primer pairProduct of target gene in R478bPrimer sequencePCR gene name (position of amplicon in R478b sequence)
10FwHtdV, putative membrane protein5′-AATCGCCTGAATCAGCTGG-3′smr10-11 (6655-7713)
11RvHtdT, putative plasmid transfer protein5′-TTCTTTACTACACCAGAGCC-3′
17FwSMR0017, putative lipoprotein5′-AACTCTTGAAAATCGTGG-3′smr17-18 (18211-19101)
18Rv5′-CTTCAGGCTATCGTTTCG-3′
TerFwTellurium resistance protein5′-ATGCAGGCTCAAGGAATCGC-3′terF (80270-81163)
TerRv5′-TTCATCGATCCACGGTCTG-3′
92FwSMR0092, hypothetical protein5′-CTATGTAAGCAATGATCCTC-3′smr92-93 (88861-89862)
93RvSMR0093, putative inner membrane protein5′-TATAGAGAGCACCGAAGG-3′
TnsDAFwTn7-like transposition protein5′-AATCCTTGTTCAGCCGG-3′tnsD (119360-120825)
TnsDARv5′-CAAAAGCCAGCCATGCCC-3′
136AfwSMR0136, hypothetical protein5′-TACGAAAATGAATTGTGGC-3′smr136 (120906-121768)
136Arv5′-AATTTACAATCTGCAGCCC-3′
ArsBFwArsenical pump membrane protein5′-AGTGAAAGACAGACGAAGCG-3′arsB (159735-160870)
ArsBRv5′-GGCAGATAGTGTGGAATGCG-3′
201FwSMR0201 hypothetical protein5′-TGTCAGGCTAAGTCACTGG-3′smr201 (180398-181466)
201Rv5′-ATTATACGGTAGATCC-3′
207FwSMR0207, conserved hypothetical protein5′-TTTCCCAAATAGGCGACGC-3′smr207-208 (190238-191131)
208RvSMR0208, hypothetical protein5′-ATGTGAAATTACTATACCGG-3′
239FwSMR0239, hypothetical protein5′-TGGAACGCGTGGTATGTGG-3′smr239-240 (219372-220364)
240RvSMR0240, hypothetical protein5′-ATACCTGCCGTTTACCC-3′
O1R_160FwO1R_160, Tn10 insertion site5′-TTATGATGCTGGGCGTACC-3′O1R_160 (202719-212075)c
O1R_160Rv5′-CACCATTACAATCACCTCC-3′
  • a The DNA from the reference plasmid R478 of the incompatibility group IncHI2 was used as control for PCR assays. Amplification conditions were initial denaturation at 94°C for 5 min and 30 cycles at 94°C for 1 min, annealing at 60°C for 30 s, and elongation at 72°C for 1 min.

  • b EMBL accession no. BX664015.

  • c Positions 183121 to 182957 in EMBL accession no. DQ517526.