Figure 1

Generation of HDGF ^Tyr transgenic mice. A: Schematic map of the Tyrosinase-HDGFgene construct. The Tyrosinase Promotor/Enhancer element is located approximately 6.1 kb in front of exonI. The HDGF gene consists of the complete exon/intron structure, except intronI was shortened from 6, 377 bp to 786 bp. The construct was separated from the vector backbone by incubation with the restriction enzyme BssHII, purified, and injected into the pronucleus of fertilized mouse egg cells. Boxes 1-6 indicate exons (white parts represent non-coding, black parts indicate coding regions). Arrows show the site of primer annealing for PCR-genotyping. The probe used for Southern blot analysis spans exonIV, intronIV and exonV. B: Examples of a polymerase chain reaction (PCR) genotyping of mouse tail DNA. Primers a and b were used to amplify the 943 bp fragment. Wildtype DNA was used as a negative control. 10 ng of Tyr-HDGFgene construct plasmid DNA was used as a positive control. C: Exemplary illustration of a Southern blot analysis of mouse tail DNA from the PCR-positive offspring 7159 and 889 and the PCR-negative offspring 7165 and wildtype mouse tail DNA as a control. The radioactively labeled probe spanning exonIV, intronIV and exonV hybridizes to both fragments of the wildtype HDGF-allele (4, 386 bp) and the Tyr-HDGFgene construct (2, 570 bp).