Plasmids or E. coli strains or primers | Relevant properties or genetic marker | Source |
---|---|---|
Plasmids | ||
pMD18-T | ColE1 ori, Ampr | Takara |
pQE31 | ColE1 ori, T5 promoter, Ampr | Qiagen |
pETDuet-1 | f1 ori, T7 promoter, Ampr | Novagen |
pQ-4CL1 | pQE31 carrying 4CL1 from P. tomentosa | This study |
pQ-CCR | pQE31 carrying CCR from P. tomentosa | This study |
pQ-4CL1–CCR | pQE31 carrying fusion gene 4CL1–CCR from P. tomentosa | This study |
pE-4CL1+CCR | pETDuet-1 carrying 4CL1 and CCR from P. tomentosa | This study |
pE-4CL1–CCR | pETDuet-1 carrying fusion gene 4CL1–CCR from P. tomentosa | This study |
Strains | ||
DH5α | Cloning host | Qiagen |
M15 (pREP4) | Expression host | Qiagen |
BL21(DE3) | Expression host | Novagen |
M-4CL1 | M15 carrying pQ-4CL1 | This study |
M-CCR | M15 carrying pQ-CCR | This study |
M-4CL1–CCR | M15 carrying pQ-4CL1–CCR | This study |
B-4CL1+CCR | BL21(DE3) carrying pE-4CL1+CCR | This study |
B-4CL1–CCR | BL21(DE3) carrying pE-4CL1–CCR | This study |
Primers | ||
pQ-4CL1-F | cgcaatggacgccacaatgaat | |
pQ-4CL1-R | actgtcttacgttgggtacg | |
pQ-CCR-F | atgccggttgatgcttcatc | |
pQ-CCR-R | gaggccttattgaatcttc | |
pQ-4CL1–CCR-F | ggcatgcatgaatccacaagaagaattc (SphI site is underlined) | |
pQ-4CL1–CCR-R | gatgaagcatcaaccggcatagaaccaccaccaccagaaccaccaccaccagaaccaccaccacctatgcctgccaacttttctttcag | |
pQ-4CL1–CCR-F | atgccggttgatgcttcatcactttcag | |
pQ-4CL1–CCR-R | gggtaccttattgaatcttcacagactcttc (KpnI site is underlined) | |
pE-4CL1-F | ttggcgcgccatgcatgaatccacaagaagaa (AscI site is underlined) | |
pE-4CL1-R | cttaagaattatgcctgccaacttttctttc (AflII site is underlined) | |
pE-CCR-F | ggggtaccatgccggttgatgcttcatcacttt (KpnI site is underlined) | |
pE-CCR-R | ccttaattaattattgaatcttcacagactcttct (PacI site is underlined) | |
pE-4CL1–CCR-F | ttggcgcgccatgcatgaatccacaagaagaa (AscI site is underlined) | |
pE-4CL1–CCR-R | cttaagaattattgaatcttcacagactcttct (AflII site is underlined) |