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Table 1 Bacterial strains and plasmids used in this study

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′