Table 1.
Bacterial strains and plasmids used in this study
| Bacterial strain or plasmid construct | Description | Reference |
|---|---|---|
| Escherichia coli XL1 Blue MRF' | Δ(mcrA) 183 Δ(mcrCB-hsdSMR-mrr) 173 endA1 supe44 thi-1 recA1 gyrA96 relA1 lac [F' proAB lacIqZΔ M15 Tn 10 (tet)] | Stratagene |
| pSE380 | Expression vector, trc promoter, lacO operator, lacI, bla | Invitrogen |
| pMRK1 | pla in pSE380 | [19] |
| pMRK4 | epo in pSE380 | [16] |
| pMRK105 | Pla with 5 aa from Epo: 161KGVRV→NQRPG | This study |
| pMRK405 | Epo with 5 aa from Pla: 159NQRPG→KGVRV | This study |
| pMRK110 | Pla with 10 aa from Epo: 161KGVRV→NQRPG, 262KN→TI, 268VSIG→ASLD | This study |
| pMRK410 | Epo with 10 aa from Pla: 159NQRPG→KGVRV, 260TI→KN, 266ASLD→VSIG | This study |
| pMRK117 | Pla with 17 aa from Epo: 35ETG→SNA, 88MNE→QNS, 155YT→SV, 161KGVRV→NQRPG, 262KN→TI, 268VSIG→ASLD | This study |
| pMRK417 | Epo with 17 aa from Pla: 33SNA→ETG, 86QNS→MNE, 153SV→YT, 159NQRPG→KGVRV, 260TI→KN, 266ASLD→VSIG | This study |
| pMRK431 | Epo with 31 aa from Pla: 33SNA→ETG, 86QNS→MNE, 153SV→YT, 159NQRPG→KGVRV, 260TI→KN, 266ASLD→VSIG, 26KELV→HEML, 101GTDVNY→ATNVNH, I257T, H79N, D92E, S210D, S167N, D219G, E249D, G22S | This study |
| pMRK431β1 | Epo with 42 aa from Pla + the signal sequence: N-terminus from Pla until aa 45; the rest similar to pMRK431 | This study |
| pMRK4β1L1 | Epo with the N-terminus from Pla until aa 45 | This study |
| pMRK410β1L1 | pMRK410 with the N-terminus from Pla until aa 45 | This study |
| pMRK417β1L1 | pMRK417 with the N-terminus from Pla until aa 45 | This study |
| pMRK431β2 | pMRK431 with β2 from Pla | This study |
| pMRK431β4 | pMRK431 with β4 from Pla | This study |
| pMRK431β5 | pMRK431 with β5 from Pla | This study |
| pMRK431β8+9 | pMRK431 with β8 & β9 from Pla | This study |
| pMRK431β10 | pMRK431 with β10 from Pla | This study |
Bold text indicates the novel substitutions compared to previous hybrid. aa, amino acid