From: Enzymatic process optimization for the in vitro production of isoprene from mevalonate
Strain/plasmid/primer | Relevant genotype/property/sequence | Source/reference |
---|---|---|
Strains | ||
E. coli BL21(DE3) | F− ompThsdS B (r − B m − B ) gal dcm rne131 (DE3) | Invitrogen |
MVK producer | BL21(DE3)/pET-ERG12 | This study |
MVD producer | BL21(DE3)/pET-ERG8 | This study |
PMK producer | BL21(DE3)/pET-ERG19 | This study |
IDI producer | BL21(DE3)/pET-IDI | This study |
ISPS producer | BL21(DE3)/pET-ISPS | This study |
Plasmids | ||
pYJM14 | pTrcHis2B derivative carryinggenes ERG8, ERG12, ERG19 and IDI, Trc promoter, ApR | [16] |
pET-ERG12 | pET30a(+) derivative carryinggenes gene ERG12, T7 promoter, KanR | This study |
pET-ERG8 | pET30a(+) derivative carryinggenes gene ERG8, T7 promoter, KanR | This study |
pET-ERG19 | pET30a(+) derivative carryinggenes geneERG19, T7 promoter, KanR | This study |
pET-IDI | pET30a(+) derivative carryinggenes gene IDI, T7 promoter, KanR | This study |
pET-ISPS | pET30a(+) derivative carryinggenes gene ISPS, T7 promoter, KanR | This study |
pACY-ISPS | pACYDuet-1 derivative carryinggenes gene ISPS, T7 promoter, CmR | This study |
Primers | ||
ERG12_F | 5′-CCCAAGCTTGGTCATTACCGTTCTTAACTTC-3′ | |
ERG12_R | 5′-CCGCTCGAGTTATGAAGTCCATGGTAAAT-3′ | |
ERG8_F | 5′-CCGGAATTCTCAGAGTTGAGAGCCTTCAG-3′ | |
ERG8_R | 5′-CCGCTCGAGTTATTTATCAAGATAAGTTT-3′ | |
ERG19_F | 5′-CGCGGATCCACCGTTTACACAGCATCCGT-3′ | |
ERG19_R | 5′-CCGCTCGAGTTATTCCTTTGGTAGACCAG-3′ | |
IDI_F | 5′-CGCGGATCCACTGCCGACAACAATAGTAT-3′ | |
IDI_R | 5′- CCGCTCGAGTTATAGCATTCTATGAATTT-3′ |