Figure 1.

(A) Organisation of the PTyr/Rab27a/hGH transgenic construct. The point mutation in Rab27a is indicated by an asterisk. (B) Screening for potential transgenic mice by Southern blotting. Genomic DNA was extracted from the tail tips of offspring from a cross between a T27aT15 transgenic and a wild type mouse and subjected to Southern blotting as described under materials and methods. Mice 4 to 8 are transgenic. (C) RT-PCR quantification of transgenic mRNA. Total RNA was extracted from eyes of adult wild type, T27aT15 and T27aT17 transgenic mice and Rab27a mRNA was quantified and normalised to Hprt expression as described previously.³ (D) Expression of transgenic Rab27a proteins. Protein lysates (50 μg) from wild type, T27aT15, T27aT17, and A27aQ24³ transgenic eyes were subjected to SDS-PAGE and analysed by immunoblotting using the monoclonal anti-Rab27a antibody 4B12 as described previously.³ Calnexin was used as a loading control and was detected using a specific polyclonal antibody.³