| Bacterial strains |
| E. coli strains |
| DH5α |
F− φ80lacZ M15 endA recA hsdR(rk−mk−) supE thi gyrA relA Δ(lacZYA-argF)U169 |
42 |
| 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 |
|
