Table 1 Plasmids and strains used in this study
From: Metabolic engineering of Escherichia coli for enhanced arginine biosynthesis
Plasmid/Strain | Relevant characteristics/genotype | Source/Reference |
|---|---|---|
Plasmids | Â | Â |
pKH15 | pCA24N (clone JW2786), argA214 | ASKA- collection [26], this work |
pKH19 | pCA24N (clone JW2786), argA215 | ASKA- collection [26], this work |
pTrc99a | Amp-R, lacIq | Lab stock |
pTrcArgP216 | pTrc99a with a mutant argP216 allele | This work |
pJB044 | pBR322 derived, infA, rop- | Lab stock [27] |
pJB044argA15 | pJB044 with argA214 downstream of infA | This work |
pJB044p1argA15 | Same as pJB044argA15 but with rrsBp1 | This work |
pArgObla | Arginine bio-sensor plasmid with bla (Amp-R) under transcriptional control of argOp | This work |
pArgObla10C | Arginine bio-sensor plasmid with bla (Amp-R) under transcriptional control of argOp with mutation in RBS | This work |
pTrcArgP216 | pTrc99a with argP216 cloned under transcriptional control of trc promoter | This work |
pJB044argAO | pJB044argA15 with argO cloned downstream of argA214 | This work |
pJB044argAP | pJB044argA15 with argp216 cloned downstream of argA214 | This work |
Strains | Â | Â |
E. coli K-12 C600 | thr-,1 leuB6, thi-1, lacY1, glnV44, supE44, rfbD1, mcrA1 | Lab stock [28] |
MG1655 | ilvG-, rfb-50, rph-1 | Lab stock |
C600+ | Same as C600 but thr + , leu + | This work |
C600+∆4 | Same as C600+ but ∆adiA, ∆speC, ∆speF, ∆argR | This work |
pTrcArgP216/C600+∆4 | C600+ ∆4 with a mutant argP216 allele | This work |
JW3932 | Auxotrophic for arginine ∆argH | [29] |
SJB001 | Glutamate independent mutant of C600+∆4 with pKH15 (clone 2) | This work |
SJB003 | Glutamate independent mutant of C600+∆4 with pKH19 (clone 2) | This work |
SJB004 | Glutamate independent mutant of C600+∆4 with pKH19 (clone 4) | This work |
SJB003A | SJB003 but no plasmid | This work |
SJB005 | SJB003A but ∆argA | This work |
SJB015 | SJB005 with pJB044argA15 | This work |
SJB025 | SJB005 with pJB044p1argA15 | This work |
SJB006 | Arginine producing mutant of C600+∆4 from biosensor selection, argP216 | This work |
SJB007 | Derivative of SJB006 from second round biosensor selection | This work |
SJB009 | SJB005 with pJB044argAO | This work |
SJB010 | SJB005 with pJB044argAP | This work |