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. 2009 Aug;155(Pt 8):2664–2675. doi: 10.1099/mic.0.027714-0

Table 1.

Bacterial strains and plasmids used in this work

Plasmids Relevant genotype and/or information* Source or reference
pET28a(+) Phagemid vector (KmrlacZ′) for expression of recombinant proteins under control of strong T7 transcription and translation signals Novagen
pD6MT pET28a(+) with M. tuberculosis accD6 His-tag fusion gene, under T7 promoter control, Kmr This study
pD6MS pET28a(+) with M. smegmatis accD6 His-tag fusion gene, under T7 promoter control, Kmr This study
pD5 pET28a(+) with M. tuberculosis accD5 His-tag fusion gene, under T7 promoter control, Kmr Gago et al. (2006)
pA3 pET28a(+) with M. tuberculosis accA3 His-tag fusion gene, under T7 promoter control, Kmr Gago et al. (2006)
pUC4K Contains aphA-3 cassette that confers Kmr Vieira & Messing (1982)
pPR27 E. coli–Mycobacterium shuttle vector, oriM temps, sacB xylE Gmr Pelicic et al. (1997)
pPR27D6 pPR27 derivative carrying accD6 : : aphA-3, Gmr Kmr This study
pCG76 E. coli/Mycobacterium shuttle vector, oriM temps, Str/Spr Guilhot et al. (1994)
pCGD6 pCG76 derivative harbouring accD6 plus 500 bp, both upstream and downstream of this ORF from M. smegmatis, Str/Spr This study
pCGHD6 pCG76 derivative harbouring M. smegmatis accD6 His-tag fusion gene, under GroEL promoter control, Str/Spr This study
E. coli
DH5α E. coli K-12 F ΔlacU169 (φ80lacZΔM15) endA1 recA1 hsdR17 deoR supE44 thi-1 λ gyrA96 relA1 Hanahan (1983)
BL21λ(DE3) E. coli B FompTgraphic file with name 2664equ2.jpg (DE3) Studier & Moffatt (1986)
BL21λ(DE3) Codon Plus E. coli B FompT hsdS (graphic file with name 2664equ2.jpg) dcm+ TetRgal (DE3) endA Hte [argU ileY leuW], CmR Stratagene
M. smegmatis
mc2155 Fast-growing strain harbouring all plasmids used herein Snapper et al. (1990)
D6SCO1 mc2155 with pPR27D6 integrated into accD6 locus, Kmr This study
D6DCO2 Mutant containing an intrachromosomal allelic exchange at accD6 locus in presence of pCGD6, Kmr Str/Spr This study
MS-CG76 mc2155 harbouring pCG76, Str/Spr This study
MS-CGD6 mc2155 harbouring pCGD6, Str/Spr This study
MS-CGHD6 mc2155 harbouring pCGHD6, Str/Spr This study

*Gmr, Gentamicin resistance; Kmr, kanamycin resistance; Str, streptomycin resistance; Spr, spectinomycin resistance; Cmr, cloramphenicol resistance; temps, temperature sensitivity.