Purpose and name | Sequence (5′ to 3′) | Description |
---|---|---|
Construction of the stm0551 mutant | ||
stm0551pKD13-F | GCTCTGATGTTTCAATGCCTTCCATCAGC ATTAACTGATTCCGGGGATCCGTCGACC | Annealing Temp.: 55°C; amplicon length: 1,550 bp |
stm0551pKD13-R | GGCACAGGGTATTTTGTTAAAGGAAAGG ATAAATCCCTGTAGGCTGGAGCTGCTTCG | Â |
Cloning | ||
stm0551-F | GGATCC CATCCTGCTTTTTCCATTGCTCTAATAT | Bam HI restriction site (underlined) |
stm0551-R | GATATC ACTCACTTAACTTTTTACAAGGCTTACG | Eco RV restriction site (underlined) |
 |  | Annealing Temp.: 55°C; amplicon length: 500 bp |
Construction of the fusion protein | ||
stm0551-TOPO-F | CACCATGGTGGCACAGGGTATTTTGTTAA | Annealing Temp.: 50°C; amplicon length: 316 bp |
stm0551-TOPO-R | ATATATATCTGGTAATATGGCTGG | Â |
fimY-TOPO-F | CACCATGCGCAGCGTACCACGCAG | Annealing Temp.: 50°C; amplicon length: 727 bp |
fimY-TOPO-R | AAAAATGTCGTGGAAAGTAACGT | Â |
E49A-TOPO-F | ATCGGCTATGC GGTCCTGACGCAACTTCCG | Mutation site (underlined) |
E49A-TOPO-R | CGGAAGTTGCGTCAGGACCG CATAGCCGAT | Mutation site (underlined) |
RT-PCR analysis | ||
fimA-RT-F | ACTATTGCGAGTCTGATGTTTG | Â |
fimA-RT-R | CGTATTTCATGATAAAGGTGGC | Â |
fimZ-RT-F | ATTCGTGTGATTTGGCGT | Â |
fimZ-RT-R | ACTTATCCTGTTGACCTT | Â |
fimY-RT-F | GAGTTACTGAACCAACAGCT | Â |
fimY-RT-R | GCCGGTAAACTACACGATGA | Â |
fimW-RT-F | AAAGTGAAAGTAAAGCGG | Â |
fimW-RT-R | AAGAGATAGATAATGCCCG | Â |
stm0551-RT-F | GCCATAAATAACCTTGTTCC | Â |
stm0551-RT-R | CATTCATATCTCAACAGCGA | Â |
16 s-F | TTCCTCCAGATCTCTACGCA |  |
16 s-R | GTGGCTAATACCGCATAACG |  |