Table 2 Strains, plasmids, and primers.
Strain, plasmid, and primer | Relevant information | Source or reference |
|---|---|---|
Bacterial strains | ||
E. coli strains | ||
DH5α | F− φ80lacZ M15 endA recA hsdR(rk−mk−) supE thi gyrA relA Δ(lacZYA-argF)U169 | |
BL21(DE3) | F− ompT hsdSB(rB−mB−) gal dcm | Novagen |
JY001 | BL21(DE3) carrying pHMS01 & pHMS05 | (This study) |
JY002 | BL21(DE3) carrying pHMS01 & pHMS06 | (This study) |
Plasmids | ||
pACYC-duet1 | Co-expression vector, Cmr. 2 MCS site with T7 promoter, lac operator, RBS. P15A replicon. 10 to 12 copy number. | Novagen |
pCDF-duet1 | Co-expression vector, Specr. 2 MCS site with T7 promoter, lac operator, RBS. CloDF13 replicon. 20 to 40 copy number. | Novagen |
pRSF-duet1 | Co-expression vector, Kmr. 2 MCS site with T7 promoter, lac operator, RBS. RSF1030 replicon(NTP1). Over 100 copy number. | Novagen |
pHMS01 | pCDF- duet1, Specr, cadA (MCS1) and acn (MCS2). | (This study) |
pHMS02 | pACYC-duet1, Cmr, cadA (MCS1). | (This study) |
pHMS03 | pRSF-duet1, Kmr, cadA (MCS1). | (This study) |
pHMS04 | pACYC-duet1, Cmr, 2 cadA (MCS1,2). | (This study) |
pHMS05 | pRSF-duet1, Kmr, 2 cadA (MCS1,2). | (This study) |
pHMS06 | pRSF-duet1, Kmr, 2 cadA (MCS1,2), citT (MCS1). | (This study) |
Primers | ||
acn F | CTCTAT CATATG ATGGAGCTCACTGTGACTGA | |
acn R | CTCT GATATC TTACTTAGAAGAAGCAGCCATC | |
cadA F (MCS1) | CTCT GGATCC AATGACCAAGCAGAGCGCAG | |
cadA R (MCS1) | CTCT GAGCTC TTAAACCAGCGGGGATTTAAC | |
cadA F (MCS2) | ATCGTC CATATG ATGACCAAGACGAGCGCAG | |
cadA R (MCS2) | CTCT CTCGAG TTAAACCAGCGGGGATTTAAC | |
citT F | CTCT GAGCTC GCACTTGATAAATTTGGAAA | |
citT R | CTCTCT GCGGCCGC TTAGTTCCACATGGCGAG | |
