Figure 2

The integration of pCAG-eGFP in the Tg mouse line and mRNA expression of eGFP and the adjacent genes.

(A) The proportional map of the pCAG-eGFP transgene integration position at the D1 band of mouse chromosome 4. There are 34 kb between exon 1 of Akr1A1 and the Prdx1 gene in WT mice and the integration of the pCAG-eGFP transgene caused a ~30-kb genomic deletion, which includes exons 1–5 of the Akr1A1 gene and the non-coding region between the Akr1A1 and Prdx1 genes. The red and black boxes represent the exons of Akr1A1 and Prdx1 genes, respectively. The blue and green boxes represent the CAG promoter and eGFP of the transgene, respectively. ATG and TAG represent the start and stop codons of the Akr1A1 gene, respectively. P1, P2 and P3 represent the oligonucleotide primers that were used for the mouse genotyping by PCR. Scale bar: 1 kb. (B) The genotyping of the pCAG-eGFP mouse line by PCR. The 629 bp and 294 bp amplicons represent the transgenic allele and non-transgenic allele, respectively. (C) IHC of the AKR1A1 protein that was expressed in WT but not in Akr1A1^eGFP/eGFP mouse liver. Scale bar: 400 μm. (D–F) mRNA expression of Akr1A1, eGFP and Prdx1 in the Akr1A1^eGFP/+, Akr1A1^eGFP/eGFP and WT mouse livers. The bars show the mean ± s.e.m. of five animals per group (n = 5); data were analyzed by a one-way ANOVA with post hoc comparisons using Duncan’s new multiple range test; **represents significant differences (P < 0.01).