TABLE 2.
RP4 TrbE mutant phenotypes
| trbE allele | Mutation | Phenotypes
|
Oligonucleotide(s) usedd | |||||
|---|---|---|---|---|---|---|---|---|
| Transfer frequencya | Dpsb
|
Pilus forma-tionc | ||||||
| PRD1 | PRR1 | Pf3 | PRD1 adsorption (%)e | |||||
| pDB126f | 1.6 × 10−1 | + | + | + | 50.4 | + | ||
| trbE | 4.1 × 10−2 | + | + | + | 38.8 | + | ||
| ΔtrbE | In-frame deletion | <1 × 10−7 | − | − | − | 0.7 | − | TGCTTGGATCATCGGTATTGCTTC (20,556-20,533), CTGGAGGCAGCATGAGTTTTG (23,089-23,103) |
| trbEHN | N-terminal His6 tag | 3.6 × 10−2 | + | + | + | 40 | + | |
| trbEH484A | Near motif A | 2.7 × 10−2 | + | + | + | 12 | + | CGTTCCGGCTGAACCTGGCCGTGCGCGACCTCGG (21,976-22,010) |
| trbEG495A | Motif A | 5.8 × 10−3 | + | + | + | 14 | + | CCACACCTTTATGTTCGCGCCGACCGGCGCAGG (22,011-22,043) |
| trbEG500A | Motif A | <1 × 10−7 | − | − | − | 0.7 | − | CGGGCCGACCGGCGCAGCTAAATCGACGCACC (22,026-22,057) |
| trbEK501A | Motif A | 1.7 × 10−4 | − | (+) | (+) | 1.6 | − | CCGACCGGCGCAGGTGCATCGACGCACCTGGCG (22,030-22,062) |
| trbEK501E | Motif A | <1 × 10−7 | − | − | − | 0.6 | − | CCGACCGGCGCAGGTGCATCGACGCACCTGGCG (22,030-22,062) |
| trbES502A | Motif A | <1 × 10−7 | − | − | − | 0.9 | − | CGACCGGCGCAGGTAAAGCGACGCACCTGGCGATCC (22,031-22,062) |
| trbED642A | Motif C | <1 × 10−7 | − | − | − | 0.8 | − | GGGCCATCTGCTCGCCGCCGAAGAGGACGGC (22,455-22,485) |
| trbED642N | Motif C | <1 × 10−7 | − | − | − | 0.8 | − | GGGCCATCTGCTCAACGCCGAAGAGGACGGC (22,455-22,485) |
| trbED642N/ D694N | Motifs C and B | <1 × 10−7 | − | − | − | 1.0 | − | GGGCCATCTGCTCAACGCCGAAGAGGACGGC (22,455-22,485), GCCGTCATCATCCTGAACGAAGCCTGGTTGATGC (22,609-22,642) |
| trbED646A | Motif C | 2.9 × 10−2 | +/− | + | + | 7.8 | − | GCTCGACGCCGAAGAGGCCGGCTTGGCGCTGTCCG (22,464-22,498) |
| trbED694A | Motif B | <1 × 10−7 | − | − | − | 0.6 | − | GCCGTCATCATCCTGGCCGAAGCCTGGTTGATGC (22,609-22,642) |
| trbED694N | Motif B | <1 × 10−7 | − | − | − | 0.8 | − | GCCGTCATCATCCTGAACGAAGCCTGGTTGATGC (22,609-22,642) |
| trbEE695A | Motif B | <1 × 10−7 | − | − | − | 0.7 | − | CCGTCATCATCCTGGACGCAGCCTGGTTGATGCTCG (22,610-22,645) |
| trbEE695Q | Motif B | <1 × 10−7 | − | − | − | 0.7 | − | CCGTCATCATCCTGGACCAAGCCTGGTTGATGCTCG (22,610-22,645) |
| trbER717A | Motif D | 3.9 × 10−6 | − | (+) | (+) | 1.1 | + | GGCTCAAGGTGCTGGCTAAGGCCAACTGCC (22,679-22,708) |
| trbEQ728A | Motif D | 5.2 × 10−3 | + | + | + | 21.9g | + | GCTGATGGCAACGGCGAGCCTGTCCGACG (22,713-22,741) |
| trbEY766A | Near motif D | 4.2 × 10−2 | + | + | + | 38.7g | + | GGACACGGCGGCCCTGGCCCGCCGCATGGGCC (22,824-22,855) |
| trbEY766F | Near motif D | 6 × 10−2 | + | + | + | 46.9g | + | GGACACGGCGGCCCTGTTCCGCCGCATGGGCC (22,824-22,855) |
| trbEΔ1 | N terminus | 4.5 × 10−4 | − | − | − | 0.7 | − | ATATATATCATATGGCCCGCATCCGCGCGGTCGATG (20,608-20,629) |
| trbEΔ2 | N terminus | 1.3 × 10−2 | + | + | + | 35.8 | + | GGGAATTTCATATGGATGCCGAACTGAAACTGAAAAAGCATC (20,626-20,653) |
| trbEHNΔ2 | N terminus | 1.8 × 10−2 | + | + | + | 32 | + | GGGAATTTCATATGGATGCCGAACTGAAACTGAAAAAGCATC (20,626-20,653) |
| trbEΔ3 | N terminus | <1 × 10−7 | − | − | − | 0.8 | − | GCGGATGCGGGCAAAGAGGATGAAC (20,619-20,595), GACGCCGGCCTGGCCGATCTGC (20,662-20,683) |
| trbEΔ4 | N terminus | <1 × 10−7 | − | − | − | 0.7 | − | GGGAATTCATATGGACGGCGTAATCGTGGGCAAGAAC (20,707-20,730) |
| trbEHNΔ5 | N terminus | <1 × 10−7 | NDh | ND | ND | ND | ND | TATATATCATATGTTCGTCGAGCTGATGTTTGACGACG (21,043-21,070) |
| trbEHNΔ6 | N terminus | <1 × 10−7 | ND | ND | ND | ND | ND | ATATATATCATATGAAGAAGTGGCGGCAGAAGATTCGC (21,520-21,543) |
| trbEΔ7 | C terminus | 4 × 10−2 | + | + | + | 44 | + | CCCCCCGGAATTCTCATTCATCAAGGGCGAGGCCCCGGCC (23,085-23,062) |
| trbEΔ8 | C terminus | 9.3 × 10−2 | + | + | + | 38 | + | CCCCCCGGAATTCTCACCGGCCACGCAGCCATTCATCCACCC (23,067-23,042) |
| trbEΔ9 | C terminus | <1 × 10−7 | − | − | − | 0.8 | − | CCCCCCGGAATTCTCATTCATCCACCCACTGGTCGCCG (23,052-23,031) |
| trbEΔ10 | C terminus | <1 × 10−7 | − | − | − | 0.9 | − | CCCCCCGGAATTCTCACTTGATGATGGCGACGGATTCCTTGTCGG (23,013-22,985) |
| trbEΔ11 | C terminus | <1 × 10−7 | − | − | − | 0.9 | − | CCCCCCGGAATTCTCAGTCGTAGAGACGGCGGCCGTTTTC (22,944-22,921) |
Number of transconjugants per donor cell. The frequencies are average values for three independent experiments.
+, Dps positive; +/−, Dps reduced; (+), weak Dps; −, Dps negative.
+, pili found; −, no pili found.
For amino acid substitutions, only the transcriptive strand primer is shown. Nucleotides derived from the original sequence are in italics; mutagenized nucleotides are in boldface type. The RP4 nucleotide coordinates are given in parentheses according to previously published sequence data (accession no. L27758).
Relative amount of label associated with the cell pellet. 14C-labeled PRD1 was adsorbed to the cells, incubated at 24°C for 15 min, and washed twice with YT medium. Radioactivity was determined in the cell and in the supernatant fractions.
The trbE gene was encoded by plasmid pDB126.
Phages were loosely bound to the cells.
ND, not determined.