Skip to main content

Advertisement

Table 3 Bacterial strains, plasmids and primers used in this study

From: Structure and functional characterization of pyruvate decarboxylase from Gluconacetobacter diazotrophicus

Strain or plasmid Genotype or description Source or reference
Strains   
G. diazotrophicus ATCC 49037 Wild type strain PAI 5 American type culture collection
E. coli DH5α F´/endA1 hsdR17 (rKmK+) supE44 thi-1 reacA1 gyrA (Nalr) relA1 Δ(lacZYA-argF)U169 (ϕ80dlacΔ(lacZ)M15) Promega Corp.
E. coli BL21-DE3 E. coli B Fdcm omp T hsd S(rB mB) gal λ(DE3) Invitrogen Corp.
Plasmids   
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   
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.
  1. Italics sections in primer sequences indicate restriction endonuclease sites.