Tetrahydrobiopterin-dependent preservation of nitric oxide–mediated endothelial function in diabetes by targeted transgenic GTP–cyclohydrolase I overexpression
J. Clin. Invest. Nicholas J. Alp, et al. 112:725 doi:10.1172/JCI17786 [Go to this article.]

Figure 1
(a) Schematic of the murine Tie2 promoter-enhancer/human GTPCH transgene. The murine Tie2 promoter and its intronic enhancer (10 kb) are depicted as gray bars, human GTPCH cDNA is depicted as a white bar, and SV40 poly A signal is depicted as a black bar. Oligonucleotide primers were used to screen genomic DNA for the presence of the transgene. Restriction endonuclease sites for SalI and XbaI are shown. GCH, GTPCH; pA, SV40 poly A; P1 and P2, oligonucleotide primers. (b) Genomic DNA analysis of potential founders. The top panel shows PCR reactions performed on DNA isolated from tail biopsies. The expected 150-bp product (filled arrowhead) was identified in founder mouse 16; linearized pTie2-GCH plasmid DNA was used as a positive control. The bottom panel shows Southern blot analysis performed to confirm the transgenic founders’ genotype and estimate transgene copy number. Founder mouse 16 showed a single hybridization fragment of 8 kb (open arrowhead), suggesting a single chromosomal integration site. The pTie2-GCH plasmid digested with XbaI was used as a positive control.