TABLE 1.
Strains, plasmids, and primers used in this study
| Strain, plasmid, or primer | Genotype | Source or reference |
|---|---|---|
| Strains | ||
| E. coli DH5α | λ− ϕ80dlacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17(rK− mK−) supE44 thi-1 gyrA relA1 | Laboratory collection |
| E. coli S17 λpir | Tpr Smr recA thi pro hsdR hsdM+ RP4-2-Tc::Mu::Km Tn7 λpir | Laboratory collection |
| C. japonicus Ueda107 | Wild type | Laboratory collection |
| C. japonicus gspD KO | Ueda107 gspD::pK18gspDKO kan+ | 11 |
| C. japonicus Δgsp | Ueda107 Δgsp | This study |
| C. japonicus Δcel5B | Ueda107 Δcel5Ba | This study |
| C. japonicus Δcel6A | Ueda107 Δcel6Ab | This study |
| C. japonicus Δcbp2E | Ueda107 Δcbp2Ec | This study |
| C. japonicus Δcbp2D | Ueda107 Δcbp2Dd | This study |
| C. japonicus Δcel5B Δcel6A | Ueda107 Δcel5B Δcel6A | This study |
| Plasmids | ||
| pRK2013 | ColE1 RK2-Mob+ RK2-Tra+; Kmr | 21 |
| pK18mobsacB | pMB1 ori mob+ sacB+; Kmr | 18 |
| pK18gspDKO | Contains 500-bp internal gspD cloned into plasmid pK18mobsacB; Kmr | 11 |
| pK18Δgsp | Contains 500 bp upstream of the gspC start codon and 500 bp downstream of the stop codon of gspK cloned into pK18mobsacB; Kmr | This study |
| pK18Δcel5B | Contains 500 bp upstream and downstream of cel5B cloned into pK18mobsacB; Kmr | This study |
| pK18Δcel6A | Contains 500 bp upstream and downstream of cel6A cloned into pK18mobsacB; Kmr | This study |
| pK18Δcbp2E | Contains 500 bp upstream and downstream of cbp2E cloned into pK18mobsacB; Kmr | This study |
| pK18Δcbp2D | Contains 500 bp upstream and downstream of cbp2D cloned into pK18mobsacB; Kmr | This study |
| Primers | ||
| Δgsp CONF (5′) | 5′-ACGGGTAGATGGTGT-3′ | This study |
| Δgsp CONF (3′) | 5′-TAATTTCCAGCCCCAG-3′ | This study |
| Δgsp INT (5′) | 5′-TGCCGATCT TTCTGT-3′ | This study |
| Δgsp INT (3′) | 5′-GATGACTCGTAGCGAT-3′ | This study |
| Δcel5B CONF (5′) | 5′-ATCTTCACGGATCACG-3′ | This study |
| Δcel5B CONF (3′) | 5′-TCCATAATGTCGATCTCG-3′ | This study |
| Δcel5B INT (5′) | 5′-AGTATTACAACGCCCA-3′ | This study |
| Δcel5B INT (3′) | 5′-GCCATTGGCATTGACT-3′ | This study |
| Δcel6A CONF (5′) | 5′-GTTGGAGTGTTAGCTG-3′ | This study |
| Δcel6A CONF (3′) | 5′-TGCGGTTGTGGTAGA-3′ | This study |
| Δcel6A INT (5′) | 5′-ATCAAGCAGCTCTGTTC-3′ | This study |
| Δcel6A INT (3′) | 5′-CACCGATACGATCCAT-3′ | This study |
| Δcbp2E CONF (5′) | 5′-ATTGATGCAGGACCTG-3′ | This study |
| Δcbp2E CONF (3′) | 5′-GTGTCCAATTGCGTAG-3′ | This study |
| Δcbp2E INT (5′) | 5′-TACAGCCTCTGCGAT-3′ | This study |
| Δcbp2E INT (3′) | 5′-GCTGATGTTATTCGCCA-3′ | This study |
| Δcbp2D CONF (5′) | 5′-CAGACGGTTGAATTTGGA-3′ | This study |
| Δcbp2D CONF (3′) | 5′-TCTTCGATTTCACGAATC-3′ | This study |
| Δcbp2D INT (5′) | 5′-TAACCTGATCCTGCAC-3′ | This study |
| Δcbp2D INT (3′) | 5′-GATGATGAGCTGCCT-3′ | This study |
a
BioCyc accession number CJA_2983.
b
BioCyc accession number CJA_2473.
c
BioCyc accession number CJA_2615.
d
BioCyc accession number CJA_2616.