|
E. coli Strains
|
|
|
| BL21 (DE3) |
E.coli B, F
-
ampT lon, with a λ prophage carrying the T7 RNA polymerase |
Novagen |
| NovaBlue |
E. coli K-12, recA endA, lacIq,lacY
|
Novagen |
|
Saccharomyces cerevisiae strains
|
|
|
| BY4741 |
MATa; his3Δ 1; leu2Δ 0; met15Δ 0; ura3Δ 0
|
This study |
| BY4741—LEU |
BY4741 + pESC-LEU vector, leu2 selection |
This study |
| BY4741—LEU-VopK |
BY4741 + pESC-LEU-VopK, LEU2 selection |
This study |
| BY4741—LEU-VopKS314A
|
BY4741 + pESC-LEU-VopKS314A, LEU2 selection |
This study |
| BY4741—LEU-VopK H353A
|
BY4741 + pESC-LEU-VopKH353A, LEU2 selection |
This study |
| BY4741—LEU-VopK E357A
|
BY4741 + pESC-LEU-VopKE357A, LEU2 selection |
This study |
| BY4741—H10 |
BY4741 + pGMH10 vector, HIS3 selection |
This study |
| BY4741—H10—VopK |
BY4741 + pGMH10—VopK, HIS3 selection |
This study |
| BY4741—H10—VopKS314A
|
BY4741 + pGMH10—VopKS314A, HIS3 selection |
This study |
| BY4741—H10—VopKH353A
|
BY4741 + pGMH10—VopKH353A, HIS3 selection |
This study |
| BY4741—H10—VopKE357A
|
BY4741 + pGMH10—VopKE357A, HIS3 selection |
This study |
| BY4741—H10—VopKS354A
|
BY4741 + pGMH10—VopKS354A, HIS3 selection |
This study |
|
Plasmids
|
|
|
| pESC-LEU |
LEU2 selection |
Agilent Technologies |
| pGMH10 |
HIS3 selection |
RIKEN |
| pET15b |
Apr, N-terminal 6His-tag expression vector |
Novagen |
| pESC-LEU-VopK |
1.275 kb VopK gene fragment amplified from genomic DNA with XhoI—NheI VopK primers by PCR and cloned into pESC—LEU which was digested with same restriction enzymes |
This study |
| pGMH10-VopK |
1.275 kb VopK gene fragment amplified from genomic DNA with BamHI-SmaI VopK primers by PCR and cloned into pGMH10 which was digested with same restriction enzymes |
This study |
| pESC-LEU-VopKS314A
|
VopK harboring A in place of S at position 314 |
This study |
| pESCL-EU-VopK H353A
|
VopK harboring A in place of H at position 353 |
This study |
| pESC-LEU-VopK E357A
|
VopK harboring A in place of E at position 357 |
This study |
| pGMH10-VopKS314A
|
VopK harboring A in place of S at position 314 |
This study |
| pGMH10-VopKH353A
|
VopK harboring A in place of H at position 353 |
This study |
| pGMH10-VopKE357A
|
VopK harboring A in place of E at position 357 |
This study |
| pGMH10-VopKS354A
|
VopK harboring A in place of S at position 354 |
This study |
