Table 1.
Bacterial strains | ||
JM109 | e14–(McrA–) recA1 endA1 gyrA96 thi-1 hsdR17 ( ) supE44 relA1 Δ(lac-proAB) [F' traD36 proAB lacIqZΔM15] | Laboratory stock |
BL21 star™(DE3) | F– ompT hsdSB( ) gal dcm rne131 | Invitrogen |
Plasmidsa | ||
pIVEX2.3d | Ampicillin resistant, C-terminal His-tag | Roche Applied Sciences |
pK7hEPO | EPO (501 bp) cloned into NdeI/SalI sites | (18) |
pK7pelBss-EPO | pK7EPO derivative containing the signal peptide sequence of pelB (66 bp) | This study |
pK7ompAss-EPO | pK7EPO derivative containing the signal peptide sequence of ompA (63 bp) | This study |
pK7phoAss-EPO | pK7EPO derivative containing the signal peptide sequence of phoA (63 bp) | This study |
pK7malEss-EPO | pK7EPO derivative containing the signal peptide sequence of malE (78 bp) | This study |
pK7ompCss-EPO | pK7EPO derivative containing the signal peptide sequence of ompC (63 bp) | This study |
pK7ompTss-EPO | pK7EPO derivative containing the signal peptide sequence of ompT (60 bp) | This study |
pK7heposs-EPO | pK7EPO derivative containing the signal peptide sequence of human EPO (81 bp) | This study |
aAll the pK7 plasmids have kanamycin resistance and the same 5′-UTR sequences including T7 promoter and RBS.