Table 1.
Bacterial strains, plasmids, and oligonucleotides used in this study
| Strain, plasmid, or oligonucleotide | Description or sequence (5′–3′) | Reference or source |
|---|---|---|
| Strains | ||
| SL1344 | Wild-type S. Typhimurium, fimbriate | 17 |
| SL1344ΔfimA | SL1344 fimA deletion mutant, nonfimbriate | This study |
| SL1344ΔfimA | SL1344 fimA partial deletion mutant, C-terminal half of gene intact, nonfimbriate | This study |
| SL1344fimA::TT | SL1344 fimA insertion mutant, nonfimbriate | This study |
| SL1344fimA::TT::fimA | SL1344 fimA insertion mutant, fimA integrated in single copy, fimbriate | This study |
| SL1344ΔfimI | SL1344 fimI deletion mutant, fimbriate | This study |
| SL1344ΔfimH | SL1344 fimH deletion mutant, nonfimbriate | This study |
| SL1344ΔfimH::fimH | SL1344 fimH deletion mutant, fimH integrated in single copy, fimbriate | This study |
| SL1344ΔfimF | SL1344 fimF deletion mutant, nonfimbriate | This study |
| SL1344ΔfimHF | SL1344 fimHF double deletion mutant, nonfimbriate | This study |
| NEB 5-α | E. coli subcloning strain | NEB (Ipswich, MA) |
| Plasmids | ||
| pGEM-T Easy | Ampr subcloning vector | Promega (Madison, WI) |
| pfimA | Ampr vector carrying fimA | This study |
| pfimAI | Ampr vector carrying fimAI | This study |
| pfimAIC | Ampr vector carrying fimAIC | This study |
| pfimAICDHF | Ampr vector carrying fimAICDHF | This study |
| pISF101 | Camr vector carrying 12.8-kbp S. Typhimurium fim gene cluster | 8 |
| pfimH | Ampr vector carrying fimH | This study |
| pfimF | Ampr vector carrying fimF | This study |
| pfimHF | Ampr vector carrying fimHF | This study |
| pLA2 | Kanr CRIM integration vector | 16 |
| pINT | Ampr CRIM integration helper plasmid | 16 |
| pLA2fimA | Kanr CRIM integration vector carrying fimA inserted at SphI/KpnI | 16 |
| pKD3 | Camr Red recombinase template plasmid | 9 |
| pKD46 | Ampr Red recombinase expression plasmid | 9 |
| Oligonucleotides | ||
| SAZ12 | ATGATCCTTCGGCGCGTTTTCATCGCTATCGGTTGTGTTTTGTTCAGCCCTGTGTAGGCTGGAGCTGCTTC | |
| SAZ13 | CTAATTGTAATTGATCAGGAAGGTCGCATCCGCGTTAGCAAGTCCGGGCTCATATGAATATCCTCCTTAG | |
| SAZ41 | ATGATAAGGAAAGGCGCGGCGCTAGCGGGGCTTGTTTTGATGTCGCCCGTTGTGTAGGCTGGAGCTGCTTC | |
| SAZ42 | TCATGGGTAAACCAGCGTAAACCACACTTCTGAATGAATTCGTCCCGGCGCATATGAATATCCTCCTTAG | |
| SAZ43 | GTTGCGGTAGTGCTATTGTCCGCAGAGGAGACAGCCAGCAAATTAGGGTTCATATGAATATCCTCCTTAG | |
| SAZ44 | ACAGGATGCCGAAACCGGG | |
| SAZ45 | CCCCGATAGCCTCTTCCGT | |
| SAZ56 | ATGACCTCTACTATTGCGAGTCTGATGTTTGTCGCTGGCGCAGCGGTTGCG | |
| SAZ83 | GTAATTCAAGGGAAATCCATGAAACATAAATTAATGACCTCTACTATTGCTGTGTAGGCTGGAGCTGCTTC | |
| SAZ84 | CCGGAGTAGGATCAGCCGCAACCGCTGCGCCAGCGACAAACATCAGACTCCATATGAATATCCTCCTTAG | |
| SAZ90 | GGCTGTCTCCTCTGCGGAC | |
| SAZ91 | TAAACGCATTCGTGCGGT | |
| SAZ92 | ATTTGATGACCAGCAGC | |
| SAZ93 | ATACGCAGCGTGTTTTC | |
| SAZ94 | GCCGGAAGACGCGCCGAC | |
| SAZ95 | CACCCGGTAAATACCGG | |
| SAZ96 | ACGTCCGTTACCGTTTG | |
| SAZ97 | CCAACCTGATTTTTCCGGCAGCG | |
| SAZ98 | AGGCCTCAGGGCAGGCG | |
| SAZ99 | GGCGGGCAACCTTCAAGTT | |
| SAZ108 | GCGTTGCTCTCGAGGGGAGTACATTTACAATAA | |
| BD4 | CGGAGGATAGCCTGAAGCAGCCGATTA | |
| BD5 | CTTCGCCCAGAGATGAGTTGGCCTGA | |
| BD11 | TATTCGCGCCTGGCCAATCA | |
| BD12 | GGCCTTCACTCTATCGTTGAGCT | |
| pLA2fimAfwd | CTGGCATGCCATTCAGGCCGTAGGTATCA | |
| pLA2fimArev | ACAGGTACCGATAGCCTCTTTCCGTTGAG | |
| fimHdelfwd | ATGAAAATATACTCAGCGCTATTGCTGGCGGGGACCGCGCTCTTTTTCACTGTGTAGGCTGGAGCTGCTTC | |
| fimHdelrev | TTAATCATAATCGACTCGTAGATAGCCGCGCGCAGTAAACGGCCCTTCCGCATATGAATATCCTCCTTAG | |
| CRIM P1 | GGCATAATAGCAATGTAC | |
| CRIM P2 | ACTTAACGGCTGACATGG | |
| CRIM P3 | ACGAGTATCGAGATGGCA | |
| CRIM P4 | AAATATGCCCTTCGCGCA |