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Table 1 Methylobacterium Strains and Plasmids Used in This Study

From: Development of a broad-host-range sacB-based vector for unmarked allelic exchange

Strain/Plasmid Relevant Properties Reference
Strains*   
AM1-W crtI::ISphoA/hah (i.e. crtI502) strain; white [14]
C2925H CmR, SmR; New England Biolabs
CM253.1 ΔmptG [18]
CM501 Isolate of wild-type M. extorquens AM1; pink This study
CM502 Isolate of AM1-W; white "wild-type" with crtI502 allele This study
CM508 Isolate of ΔmptG strain CM253.1 This study
CM1122 KmR; ΔhprA::kan; pCM431 integrated into CM501 This study
CM1123 KmR; ΔhprA::kan crtI502; pCM431 integrated into CM502 This study
CM1203 ΔhprA; kan deleted from CM1122 This study
CM1204 ΔhprA crtI502; kan deleted from CM1123 This study
CM1263 TcR, sucroseS; pCM441 integrated into CM501; crtI502 upstream; white This study
CM1264 TcR, sucroseS; pCM441 integrated into CM501; crtI upstream; pink This study
CM1265 TcR, sucroseS; pCM440 integrated into CM502; crtI upstream; pink This study
CM1266 TcR, sucroseS; pCM440 integrated into CM502; crtI502 upstream; white This study
M. extorquens AM1 RifR isolate; pink [9]
Plasmids   
pCM157 TcR; broad-host-range cre expression vector [4]
pCM184 ApR, KmR, TcR; broad-host range cre-lox allelic exchange vector [4]
pCM411 ApR, KmR; mptG region cloned into pCR2.1 This study
pCM417 ApR, KmR; crtI region cloned into pCR2.1 This study
pCM424 ApR, KmR; ΔmptG region cloned into pCR2.1 This study
pCM426 ApR, KmR; crtI502 region cloned into pCR2.1 This study
pCM428 ApR, KmR; pCR2.1 with hprA upstream flank This study
pCM429 ApR, KmR; pCR2.1 with hprA downstream flank This study
pCM430 ApR, KmR, TcR; pCM184 with hprA upstream flank from pCM428 This study
pCM431 ApR, KmR, TcR; pCM430 with hprA downstream flank from pCM429 This study
pCM432 ApR, TcR; synthetic linker introduced into pCM184 This study
pCM433 ApR, CmR, TcR; pCM432 with sacB-cat fragment from pDS132; broad-host-range sacB-based allelic exchange vector This study
pCM433r ApR, CmR, TcR; pCM433 with sacB-cat fragment in the opposite orientation as in pCM433 This study
pCM434 ApR, KmR; hprA region cloned into pCR2.1 This study
pCM435 ApR, CmR, TcR; hprA region from pCM434 cloned into pCM433 This study
pCM436 ApR, CmR, TcR; mptG region from pCM411 cloned into pCM433 This study
pCM437 ApR, CmR, TcR; ΔmptG region from pCM424 cloned into pCM433 This study
pCM438 ApR, KmR; ΔhprA region cloned into pCR2.1 This study
pCM439 ApR, CmR, TcR; ΔhprA region from pCM438 cloned into pCM433 This study
pCM440 ApR, CmR, TcR; crtI region from pCM417 cloned into pCM433 This study
pCM441 ApR, CmR, TcR; crtI502 region from pCM426 cloned into pCM433 This study
pCR2.1 ApR, KmR; PCR cloning vector Invitrogen
pDS132 ApR, CmR;, sacB-based allelic exchange vector with λpir-dependent R6Kori [10]
pRK2073 SmR; helper plasmid supplying IncP tra functions [17]
  1. *All M. extorquens AM1 strains are also RifR [9]. Antibiotic resistances are indicated as follows, Ap (ampicillin), Cm (chloramphenicol), Km (kanamycin), Rif (rifamycin), Sm (streptomycin), Tc (tetracycline).