Table 1.
Strains and plasmids used in this study
| Strain/plasmid | Relevant features | Reference/source |
|---|---|---|
| Strains | ||
| E. coli Top10 | F- mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 deoR recA1 araD139 Δ(ara-leu)7697 galU galK rpsL (StrR) endA1 nupG | Invitrogen |
| C. acetobutylicum
ATCC 824 (COSMIC) |
CRG1268 strain used in the ERANET (http://www.sysmo.net) SysMO project COSMIC. Originally from G. Bennet, RICE University, USA | H Bahl University of Rostock |
| C. acetobutylicum
ATCC 824 |
CRG3286, resourced from the ATCC in November 2011 | ATCC |
| C. acetobutylicum
CRG1545 |
ΔpyrE mutant of CRG1268 made using the ACE vector pMTL-JH12 | [24] |
| C. acetobutylicum
CRG3899 |
ΔpyrE mutant of CRG3286 equivalent to CRG1268 | This study |
| C. acetobutylicum
CRG3520 |
Δspo0A of CRG1545 (ΔpyrE) made using pMTL-ME3::spo0A | This study |
| C. acetobutylicum
CRG4889 |
Δspo0A mutant of CRG1545 generated from CRG3520 by restoration of the pyrE allele to wild-type using pMTL-ME6 | This study |
| C. acetobutylicum
CRG4890 |
CRG3520 (Δspo0A) carrying a functional copy of spo0A and its own promoter inserted into the genome using the ACE vector pMTL-ME6X::spo0A concomitant with restoration of the pyrE allele | This study |
| C. acetobutylicum CRG4891 | CRG3520 (Δspo0A) carrying a functional copy of spo0A under the control of the fdx promoter inserted into the genome using the ACE vector pMTL-ME6X::spo0A concomitant with restoration of pyrE | This study |
| C. acetobutylicum
CRG4893 |
In-frame deletion mutant of amyP (CaP0168) made using pMTL-ME3::amyP (pyrE vector with amyP KO cassette) | This study |
| C. acetobutylicum
CRG4894 |
restoration of the pyrE allele to wild-type using pMTL-ME6 | This study |
| C. acetobutylicum
CRG4896 |
CRG4893 (ΔamyP) carrying a functional copy of amyP and its own promoter inserted into the genome using the ACE vector pMTL-ME6C::amyP concomitant with restoration of the pyrE allele | This study |
| C. acetobutylicum
CRG4897 |
CRG4894 (ΔamyP) carrying a functional copy of amyP under the control of the fdx promoter inserted into the genome using the ACE vector pMTL-ME6X::amyP concomitant with restoration of pyrE allele | This study |
| C. acetobutylicum
CRG4883 |
In-frame deletion mutant of glgA (Cac2239) made using pMTL-ME3::glgA (pyrE vector with glgA KO cassette) | This study |
| C. acetobutylicum
CRG4898 |
ΔglgA mutant of CRG4883 generated from CRG1545 by restoration of the pyrE allele to wild-type using pMTL-ME6 | This study |
| C. acetobutylicum
CRG4901 |
CRG4883 (ΔglgA) carrying a functional copy of glgA and its own promoter inserted into the genome using the ACE vector pMTL-ME6C::glgA concomitant with restoration of the pyrE allele | This study |
| C. acetobutylicum
CRG4902 |
CRG4883 (ΔglgA) carrying a functional copy of glgA under the control of the fdx promoter inserted into the genome using the ACE vector pMTL-ME6X::glgA concomitant with restoration of pyrE allele | This study |
| C. acetobutylicum CRG2404 | In-frame deletion mutant of spo0A gene made using pMTL-SC7515::spo0A | This study |
| C. acetobutylicum CRG2403 | In-frame deletion mutant of the type II Restriction system Cac1502 made using pMTL-SC7515::cac824I | This study |
| Plasmids | ||
| pAN-2 | E.coli vector (ϕ3T I methyltransferase gene, tet, p15a) | CRG685 [13] |
| pMTL85141 | E.coli- Clostridium Shuttle vector (pIM13 catP ColEI) | CRG1000 [31] |
| pMTL-ME2 | Clostridium KO vector (pCB102 catP ColE1 pyrE) | CRG1668 [25] |
| pMTL-ME3 | Clostridium KO vector (pIM13 catP ColE1 pyrE) | CRG1671This study |
| pMTLSC7315 | Clostridium KO vector (pCB102 catP ColE1 traJ codA) | CRG1424 [21] |
| pMTLSC7515 | Clostridium KO vector (pIM13 catP ColE1 traJ codA) | CRG1303 This study |
| pMTL-ME6 | C. acetobutylicum ACE correction vector (pIM13 catP ColE1 `pyrE) | CRG1861 This study |
| pMTL-ME6C | C. acetobutylicum ACE complementation vector (pIM13 catP ColE1 pyrE lacZ’ MCS) | CRG2483 [25] |
| pMTL-ME6X | C. acetobutylicum ACE expression vector (pIM13 catP ColE1 `pyrE P fdx, lacZ’ MCS) | CRG2482 [25] |
| pMTL-ME3::spo0A | pMTL-ME3 + spo0A KO cassette (pIM13 catP ColE1 pyrE) | CRG1701 This study |
| pMTL-ME3::amyP | pMTL-ME3 + amyP KO cassette (pIM13 catP ColE1 pyrE) | CRG3727 This study |
| pMTL-ME3::glgA | pMTL-ME3 + glgA KO cassette (pIM13 catP ColE1 pyrE) | CRG4881 This study |
| pMTL-SC7515::spo0A | pMTLSC7515 + spo0A KO cassette (pIM13 catP ColE1 codA) | CRG1672 This study |
| pMTL-SC7515::cac824I | pMTLSC7515 + cac824I KO cassette (pIM13 catP ColE1 codA) | CRG2293 This study |
| pMTL-ME6C::spo0A | ACE complementation vector for spo0A | CRG3739 This study |
| pMTL-ME6X::spo0A | ACE overexpression vector for spo0A | CRG3738 This study |
| pMTL-ME6C::amyP | ACE complementation vector for amyP | CRG3723 This study |
| pMTL-ME6X::amyP | ACE overexpression vector for amyP | CRG3722 This study |
| pMTL-ME6C::glgA | ACE complementation vector for glgA | CRG4900 This study |
| pMTL-ME6X::glgA | ACE overexpression vector for glgA | CRG4880 This study |