Strain or plasmid | Genotype or description | Source or reference |
---|---|---|
Strains | Â | Â |
G. diazotrophicus ATCC 49037 | Wild type strain PAI 5 | American type culture collection |
E. coli DH5α | F´/endA1 hsdR17 (rK−mK+) supE44 thi-1 reacA1 gyrA (Nalr) relA1 Δ(lacZYA-argF)U169 (ϕ80dlacΔ(lacZ)M15) | Promega Corp. |
E. coli BL21-DE3 | E. coli B F−dcm omp T hsd S(rB− mB−) gal λ(DE3) | Invitrogen Corp. |
Plasmids | Â | Â |
pGEM-T | Apr; T-tailed PCR product cloning vector | Promega Corp. |
pET17b | Apr; ColE1 replicon, HIS-tag expression vector | Novagen Corp. |
pET28a | Kanr; ColE1 replicon, HIS-tag expression vector | Novagen Corp. |
pGD | Kanr; ColE1 replicon; G. diazatrophicus pdc gene cloned into pET28a | This study |
Primers | Â | Â |
GDPDCpETF | 5'-GGAATTCCATATG ACCTATACCGTTGGACG-3' | This study |
GDPDCpETR | 5'-CCGCTCGAG TCAGCCCGCGCGCGGC-3' | This study |
GDPDCseq | 5'-ATCGACGCGCTGCTGAGCCC-3' | This study |
T7 promoter | 5'-TAATACGACTCACTATAGGG-3' | Promega Corp. |
T7 terminator | 5’-GCTAGTTATTGCTCAGCGG-3’ | Promega Corp. |