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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 (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

  

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.