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. 2015 Jul 2;10(7):e0127670. doi: 10.1371/journal.pone.0127670

Fig 6. Scheme of the 5′-flanking region of the HIBADH gene and the identification of the core promoter region.

Fig 6

The red arrow represents the previously reported SNP (g.-165 T>C, rs 133569227). Fragments pGL3-1548 (-1,548 bp to +792), pGL3-1013 (-1,013 bp to +792), pGL3-703 (-703 bp to +792), pGL3-703→+175 (-703 bp to +175), and pGL3+175 (+175 bp to +792) were amplified by PCR to produce the reporter constructs. Each fragment with wild-type alleles was cloned into the pGL3-Basic vector and transfected into MLTC-1 cells. Relative luciferase activity of a series of truncated constructs in the HIBADH 5x-flanking region was measured by dual luciferase assays of the MLTC-1 cells. For each construct, plasmid DNA extracted from 6 to 9 colonies was used. Results are presented as the average fold of firefly luciferase activity vs. the Renilla control vector (mean x S.D., n = 6 to 9). The asterisk indicates P < 0.01 vs. the pGL3 basic control.