Table 1 Bacterial strains and plasmids used in this study

Escherichia coli XL1 Blue MRF'

Δ(mcrA) 183 Δ(mcrCB-hsdSMR-mrr) 173 endA1 supe44 thi-1 recA1 gyrA96 relA1 lac [F' proAB lacI^qZΔ 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