Figure 3

Detection of Rab27a mRNA. (Panel A) Schematic depiction of transgenic (rat) versus endogenous (mouse) Rab27a coding sequence. Eco RI or Sma I restriction sites as well as resulting fragment sizes are indicated. (Panel B) Comparison of transgenic versus endogenous Rab27a expression in the eye by RT-PCR and restriction digestion of PCR products. Rab27a mRNA from wild-type and independent mutant transgenic lines (Rab27a^T23N and Rab27a^N133I) amplified by RT-PCR was digested with Eco RI or Sma I and electrophoresed as described under "Methods". Rat Rab27a cDNA was used as a control. (Panel C) Determination of Rab27a expression (transgenic and endogenous) in eyes by RT-PCR (+). The oligonucleotides and conditions were as described under "Methods". The PCR products were analysed on an agarose gel stained with ethidium bromide. Reactions performed without reverse transcriptase are indicated (-). The arrows on the left-hand side indicate the positions of DNA marker sizes. Hprt expression was used as an internal control.