Table 1 Bacterial strains and plasmids used in this study
Strain or plasmid | Relevant characteristics | Source or reference |
|---|---|---|
Strains | ||
E. coli AAEC072A | MG1655 ΔfimA-H | [44] |
E. coli MKS12 | MG1655 ΔfimA-H ΔfliCD | [21] |
E. coli MKS12P lac peb1 | MKS12 carrying peb1 under P lac at the ΔfliC site | This study |
E. coli MKS111 | MG1655 ΔfimA-H ΔfliCD ΔflgM | This study |
E. coli MKS111ΔHBB | MG1655 ΔfimA-H ΔfliCD ΔflgM ΔfliFGH ΔflgDEFGHIJ | This study |
E. coli DH10B | endA- RecA- Dam/Dcm+ | [66] |
Plasmids | ||
pMCS3'UTR | Multiple cloning site preceding the 3'UTR sequence of fliC in Pvu I-EcoR I of pBR322 | [21] |
pPeb15'3' | 173 bp fliC promoter sequence and 702bp fragment encoding mature Peb1 in Pvu I-Sca I site of pMCS3'UTR | [21] |
pLA25 | PfliC and fragment encoding mature Peb1, named fliC 5'UTR-peb1, in BglI I-Xba I site of pMCS3'UTR | This study |
pLA26 | 3'UTR removed by digestion at Sca I-EcoR I sites of pLA25 | This study |
pLA31 | Fragment containing PfliC and truncated fliC (encoding N-terminal 183 aa) in BglI I-Xba I site of pMCS3'UTR | This study |
pLA32 | Fragment containing PfliC, truncated fliC (encoding N-terminal 20 aa) and peb1 (without ss) in BglI I-Xba I site of pLA25 | This study |
pTAR15G | Insertion of five glycines at residue D42 of Peb1. pLA25 background. | This study |
pTAR1P | Substitution of four separate polar residues in Peb1 TAR1 region with alanine. pLA25 background. | This study |
pTAR2P | Substitution of four separate polar residues in Peb1 TAR2 region with alanine pLA25 background. | This study |
pTAR1A | Substitution of seven residues in Peb1 TAR1 region with alanine. pLA25 background. | This study |
pTAR1G | Substitution of seven residues in Peb1 TAR1 region with glycine. pLA25 background. | This study |
pTAR2A | Substitution of eight residues in Peb1 TAR2 region with alanine. pLA25 background. | This study |
pTAR2G | Substitution of eight residues in Peb1 TAR2 region with glycine. pLA25 background. | This study |