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Table 1 Plasmids and bacterial strains used in this work with relevant genotype characteristics

From: Insights into bacterial CO2 metabolism revealed by the characterization of four carbonic anhydrases in Ralstonia eutropha H16

Strain or plasmid Relevant characteristics Reference or source
Ralstonia eutropha
H16 Wild-type gentamycin resistant (Genr) ATCC 17699
Re2061 H16 ∆phaCAB (Genr) [11]
Re2427 H16 ∆can This study
Re2428 H16 ∆caa This study
Re2430 H16 ∆cag This study
Re2437 H16 ∆can2 This study
Re2436 H16 ∆cancan2caacag This study
Escherichia coli
DH10-beta competent cells Strain suitable for high efficiency transformation New England Biolabs
BL21(DE3) Strain suitable for transformation and protein expression New England Biolabs
S17-1 Conjugation strain for transfer of plasmids into R. eutropha [18]
Plasmids
pBBR1MCS-2 Broad-host-range cloning vector confers kanamycin resistance (Kanr) [19]
pCan pBBR1MCS-2 containing can gene (H16 A0169) (Kanr) This work
pCan2 pBBR1MCS-2 containing can2 gene (H16 B2270) (Kanr) This work
pCaa pBBR1MCS-2 containing caa gene (H16 B2403) (Kanr) This work
pCaaB pBBR1MCS-2 containing caa gene without the N terminal signaling peptide sequence (H16 B2403) (Kanr) This work
pCag pBBR1MCS-2 containing cag gene (H16 A1192) (Kanr) This work
pETCan pET14b containing can gene (H16 A0169) (Ampr) This work
pETCan2 pET14b containing can2 gene (H16 B2270) (Ampr) This work
pETCaa pET14b containing caa gene (H16 B2403) (Ampr) This work
pETCag pET14b containing cag gene (H16 A1192) (Ampr) This work
pStrepCan pET51b containing can gene (H16 A0169) (Ampr) This work
pStrepCaa pET51b containing caa gene (H16 B2403) (Ampr) This work
pStrepCaaB pET51b containing caa gene without the N terminal signaling peptide sequence (H16 B2403) (Ampr) This work
pRARE Overcoming the codon bias of E. coli for enhanced protein expression (Camr) [20]
pJV7 pJQ200Kan with ∆phaC1 inserted into BamHI restriction site, confers kanamycin resistance (Kanr) [9]
pJV7 ∆can pJV7 with ∆phaC1 allele removed by BamHI digestion and replaced with ∆can allele (Kanr) This work
pJV7 ∆can2 pJV7 with ∆phaC1 allele removed by BamHI digestion and replaced with ∆can2 allele (Kanr) This work
pJV7 ∆caa pJV7 with ∆phaC1 allele removed by BamHI digestion and replaced with ∆caa allele (Kanr) This work
pJV7 ∆cag pJV7 with ∆phaC1 allele removed by BamHI digestion and replaced with ∆cag allele (Kanr) This work
pRFP pBBR1MCS-2 containing rfp gene amplified from JBp000066 kindly offered by J. Mueller (JBEI) (Kanr) This work
pCAA_RFP pBBR1MCS-2 containing caa gene fused by a 6aa linker to the rfp gene (Kanr) This work