Table 2

Functional analysis of the two human ODC alleles

A fragment (−462 to +3070 bp) of the human ODC gene was isolated by PCR from individuals homozygous for either A or G allele and cloned into the pGL3 plasmid (Promega) in front of a modified firefly luciferase gene. NIH 3T3 cells were transiently transfected using Lipofectin reagent with the above constructs (1 μg) along with 0.1 μg of a renilla luciferase plasmid (pRL; Promega). Luciferase activities were measured in cell extracts 36 h post-transfection. Luciferase activity in cells transfected with empty pGL3 controls was 0.12 ± 0.01 (n = 3).

ExperimentLuciferase activityaA/G ratio
A alleleG allele
139.713.32.98
238.912.13.21
328.48.63.30
422.68.92.54
522.96.83.37
Mean± SD30.5± 8.369.94± 2.683.08± 0.3
  • a Ratio of relative firefly luminescence units/μg of protein to relative renilla luminescence units/μg of protein measured in the same cell extract.