Table 1 Strains, plasmids and primers used in this study
From: Engineering E. colistrain for conversion of short chain fatty acids to bioalcohols
Name | Description | Reference or source |
|---|---|---|
Strains | ||
Clostridium acetobutylicum | ATCC #824 | |
E. coli MG1655 | F- LAM- rph-1 | CGSC #6300 |
E. coli B | F- | CGSC #5713 |
E. coli DH5α | F- Φ80lacZΔM15 Δ(lacZYA-argF) U169 recA1 endA1 hsdR17 (rK–, mK+) phoA supE44 λ– thi-1 gyrA96 relA1 | Invitrogen |
E. coli M15 | F- thi lac mtl, pREP4 plasmid | Qiagen |
E. coli BW25113 | rrnB DElacZ4787 HsdR514 DE(araBAD)567 DE(rhaBAD)568 rph-1 | CGSC #7636 |
E. coli ΔfadD | BW25113, ΔfadD :: FRT-kan-FRT | CGSC #9503 |
E. coli ΔadhE | BW25113, Δadhe ::FRT-kan-FRT | CGSC #9113 |
E. coli ΔfrdA | BW25113, ΔfrdA :: FRT-kan-FRT | CGSC #10964 |
Plasmids | ||
pQE30 | bla, cloning vector | Qiagen |
pQE-adhE2 | pQE30 with adhe2 gene from C. acetobutylicum cloned between BamHI and SalI sites | This study |
pQE-ptb/buk | pQE30 with ptb-buk operon from C. acetobutylicum cloned between SalI and PstI sites | This study |
pQE-adhE2/ptb/buk | pQE-adhE2 with ptb-buk operon from C. acetobutylicum cloned between SalI and PstI sites | This study |
Primers* | ||
P1 | ATCGGATCCATGAAAGTTACAAATCAAAAA | This study |
P2 | ACTGGTCGACTTAGTGGTGGTGGTGGTGGTGAAATGATTTTATATAGATATC | This study |
P3 | ACTGGTCGACGAAGGAGATATACCATGATTAAGAGTTTTAATGAAAT | This study |
P4 | GTCTGCAGTTAGTGGTGGTGGTGGTGGTGTTTGTATTCCTTAGCTTTTTC | This study |