TABLE 3.
TrbC mutant phenotypes
| trbC allele | Phenotype
|
Primer usede (bp coordinates) | |||||
|---|---|---|---|---|---|---|---|
| TrbC maturationa | Transfer frequencyb | Dpsc
|
Pilus formationd | ||||
| PRD1 | PRR1 | Pf3 | |||||
| Wild type | C | 0.9 × 10−1 | + | + | + | + (b) | GAGCTCGGTACCCGGGGATCC, ATGGAAGCTTGATTAGGCGAG CCGTCCAGCCGC (20,214–20,238) |
| mtrbC45 | NA | <1 × 10−7 | − | − | − | − | Reference 20 |
| ΔtrbC | NA | <1 × 10−7 | − | − | − | − | GTATTTCCAATGACAACGGCGGTACCGTTC (19,788–19,801), GAGC CGTCCAGCCGGTACCGCACGCACGGC (20,199–20,228) |
| trbCS37A | C | 0.9 × 10−1 | + | + | + | + (b) | GGCGATGGCCGCGGAAGGCACC (19,895–19,916) |
| trbCS37C | C/L | 0.9 × 10−1 | − | − | − | − | GGCGATGGCCTGCGAAGGCACC (19,895–19,916) |
| trbCS37G | C | 0.9 × 10−1 | + | + | + | + (b) | GGCGATGGCCGGGGAAGGCACC (19,895–19,916) |
| trbCS37P | NAf | <1 × 10−7 | − | − | − | − | GGCGATGGCCCCGGAAGGCACC (19,895–19,916) |
| trbCS37T | C | 0.8 × 10−1 | + | + | + | + (b) | GGCGATGGCCACGGAAGGCACC (19,895–19,916) |
| trbCE38Q | C | 0.9 × 10−1 | + | + | + | + (b) | GATGGCCTCGCAAGGCACCGGC (19,898–19,919) |
| trbCG42E | C | <1 × 10−7 | − | − | − | − | AGGCACCGGCGAAAGCTTGCC (19,911–19,934) |
| trbCE47Q | C | <1 × 10−7 | − | − | − | − | CTTGCCATATCAGAGCTGGCTG (19,926–19,948) |
| trbCG59R | C | <1 × 10−7 | − | − | − | − | CTCCGTAACCCGCCCGGTGGCC (19,961–19,982) |
| trbCI78S | C | <1 × 10−7 | − | − | − | − | Reference 18 |
| trbCF79Δ | C | <1 × 10−7 | − | − | − | − | CGGCGTGCTGATCΔGGCGGCGAACTCA (20,018–20,046) |
| trbCE82Q | C | 0.7 × 10−1 | + | + | + | + (b) | CTTCGGCGGCCAACTCAACGCC (20,030–20,051) |
| trbCV96G | C | <1 × 10−7 | − | − | − | − | CCTGGTTCTGGGCATGGCGCTG (20,072–20,096) |
| trbCV106M | C | 0.9 × 10−1 | + | + | + | + (b) | CGCGCAGAACATGATGAGCACC (20,102–20,123) |
| trbCF110A | C | 0.9 × 10−3 | − | − | − | − | GATGAGCACCGCCTTCGGTCGTG (20,117–20,138) |
| trbCF110Y | C | <1 × 10−7 | − | − | − | − | GATGAGCACCTACTTCGGTCGTG (20,117–20,138) |
| trbCF111A | C | <1 × 10−7 | + | + | + | − | GATGAGCACCTTCGCCGGTCGTG (20,117–20,138) |
| trbCF111Y | C | 0.3 × 10−7 | + | + | + | − | GATGAGCACCTTCTACGGTCGTG (20,117–20,138) |
| trbCG112A | C | 0.3 × 10−7 | + | + | + | + (b) | CACCTTCTTCGCTCGTGGTGCC (20,120–20,141) |
| trbCG112D | L | <1 × 10−7 | − | − | − | − | Reference 18 |
| trbCG112S | C | 0.8 × 10−1 | + | + | + | + (b) | CACCTTCTTCAGTCGTGGTGCC (20,120–20,141) |
| trbCR113A | C | 0.9 × 10−1 | + | − | − | − | CTTCTTCGGTGCTGGTGCCGAA (20,123–20,144) |
| trbCR113K | C | 0.9 × 10−1 | + | − | − | + (b) | CTTCTTCGGTAAGGGTGCCGAA (20,123–20,144) |
| trbCG114A | C | <1 × 10−7 | + | + | + | + (b) | CTTCGGTCGTGCGGCCGAAATC (20,126–20,147) |
| trbCG114C | L | <1 × 10−7 | − | − | − | − | CTTCGGTCGTTGTGCCGAAATC (20,126–20,147) |
| trbCG114L | L | <1 × 10−7 | − | − | − | − | CTTCGGTCGTCTTGCCGAAATC (20,126–20,147) |
| trbCG114S | C/L | 0.9 × 10−1 | + | − | − | + (b) | CTTCGGTCGTAGTGCCGAAATC (20,126–20,147) |
| trbCG114T | L | <1 × 10−7 | − | − | − | − | CTTCGGTCGTACTGCCGAAATC (20,126–20,147) |
| trbCA115G | C/L | 0.7 × 10−1 | − | − | − | + (i) | GGTCGTGGTGGCGAAATCGCGG (20,129–20,150) |
| trbCA115T | C | 0.9 × 10−1 | + | + | + | + (b) | GGTCGTGGTACCGAAATCGCGG (20,129–20,150) |
| trbCE116Q | L | <1 × 10−7 | − | − | − | − | CGTGGTGCCCAAATCGCGGCC (20,132–20,153) |
| trbCI117H | C | 0.8 × 10−1 | NA | NA | NA | NA | GAGCTCGGTACCCGGGGATCC, ATGGAAGCTTGATTAGTGTTC GGCACCACGACC (20,130–20,144) |
| trbCΔ3 | C | NA | NA | NA | NA | NA | GAGCTCGGTACCCGGGGATCC, ATGGAAGCTTGATTACGCGAT TTCGGCACC (20,136–20,150) |
| trbCΔ3.05 | C | NA | NA | NA | NA | NA | GAGCTCGGTACCCGGGGATCC, ATGGAAGCTTGATTAGATTTC GGCACCACG (20,133–20,147) |
| trbCΔ3.1 | L | NA | NA | NA | NA | NA | GAGCTCGGTACCCGGGGATCC, ATGGAAGCTTGATTAGATTTC GGCACCACG (20,130–20,144) |
| trbCΔ4 | L | NA | NA | NA | NA | NA | GAGCTCGGTACCCGGGGATCC, ATGGAAGCTTGATTAACCACG ACCGAAGAA (20,124–20,138) |
Detection by Western blotting and MS as described in Materials and Methods. Pilin maturation is characterized as follows: C, circular pilin; L, linear TrbC*; C/L, circular pilin and linear TrbC* were detectable simultaneously; NA, not applicable.
Transconjugants per donor cell after 1 h of incubation with recipient HB101 Nxr at 37°C as described in Materials and Methods. The given frequencies represent the average values of three independent experiments.
Donor phage specificity (Dps) is characterized as follows: +, Dps positive; −, Dps negative; NA, not applicable. Data were derived from standard phage plaque assays and electron microscopy; for details, see Materials and Methods.
Pilus formation is characterized as follows: + (b), great amounts of pili were only detectable as bundles; + (i), few pili were only detectable as individuals; −, no pili were detectable; NA, not applicable.
For point mutations, only the transcriptive strand primer is given, and for deletion mutants and the wild type, both the primers used are given. Nucleotides derived from the original sequence are written in italics; mutagenized nucleotides are in boldface letters. RP4 coordinates of the nucleotides are given in parentheses according to published sequence data (GenBank accession no. M93696).
For details, see Results.