Table 2.
Strains, plasmids, and primers used in this study
| Strains, plasmids, and primers | Description a | Source or reference |
|---|---|---|
| Strains | ||
| B. velezensis sd | Source of carCby gene | This lab |
| E. coli DH5α | clone host | This lab |
| E. coli BL21(DE3) | Expression host | This lab |
| Plasmids | ||
| pEGFP-N3 | Containing gfp gene (720 bp), Ampr | This lab |
| pMD18-T/inak | T-clone vector ligated with the synthesized inak gene (699 bp) from Pseudomonas syringae, Ampr | This lab |
| pMD18-T/inpn | Containing N-terminal of inak gene (537 bp), Ampr | This lab |
| pET-28a( +) | Expression vector, Kanr | Novagen |
| pET-28a( +)/CarCby | Expression vector coding for carCby | This study |
| pET-28a( +)/CarCby/INPN | Expression vector coding for carCby and INPN | This study |
| pET-28a( +)/CarCby/GFP | Expression vector coding for carCby and GFP | This study |
| pET-28a( +)/CarCby/INPN/GFP | Expression vector coding for carCby, INPN and GFP | This study |
| Primers (5' → 3')a | Sequences | |
| P1 | AACTTTAAGAAGGAGATATACCatgacaaaacttaccgttcaaa | |
| P2 | TCGACGGAGCTCGAATTCGGATCTGCTTGAAACAGGATACGGCGT | |
| P3 | TCCTGTTTCAAGCAGATCCGAATATGACTCTCGACAAGGCGTTGG | |
| P4 | TGCTCGAGTGCGGCCGCAAGCTTGGTCTGCAAATTCTGCGGC | |
| P5 | TCCTGTTTCAAGCAGATCCGAATATGGTGAGCAAGGGCGAGG | |
| P6 | TGCTCGAGTGCGGCCGCAAGCTTCTTGTACAGCTCGTCCATGCC | |
| P7 | CAGAATTTGCAGACCAAGCTTATGGTGAGCAAGGGCGAGG | |
| P8 | GTGGTGGTGGTGCTCGAGCTTGTACAGCTCGTCCATGCC | |
| Ser190A-For | ATTTGGTGAAGCGGCAGGCGGCATGAGCATCG | |
| Ser190A-Rev | CTGCCGCTTCACCAAATATCGTGACGTTATCC | |
| Glu306A-For | CAATCGTGATGCAGCATATTTGTTTTTCACCCCTGA | |
| Glu306A-Rev | ATGCTGCATCACGATTGGTTCCGATGATCATA | |
| His395A-For | ATAAAGCCGTTGCCGCTCTGGAATTGCCGTTTGT | |
| His395A-Rev | AGCGGCAACGGCTTTATGAAACGGCGGTGTTT |
a The upstream and downstream recombined sequences with plasmids are underlined. The modified codons are shown in bold with red color