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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