University of Virginia Health System

I joined the Scrable lab in May 2002 as a senior lab specialist. I am known as an expert cloner and am currently constructing two targeting vectors that will be used to generate recombinant genomes in the mouse.

1. To generate novel lines of repressor mice that express the lac repressor in tissue- and developmental stage-specific patterns, I am preparing a promoter trap vector. This vector consists of a promoter-less lacI gene followed by a hygromycin-resistance/EGFP/ luciferase triple fusion coding sequence with its own translation start site (IRES). I have already demonstrated that each component of the triple fusion is functional by generating hygromycin-resistant stable cell lines that exhibit green fluorescence and measurable luminescence in the presence of the luciferase substrate, luciferin. With this vector, we can develop a library of ES cell clones with randomly trapped promoters that, when used to generate mice, will produce repressor expression patterns that depend on the expression pattern of the trapped promoter.
   
   
2. To generate a regulatable version of the endogenous p53 gene, I am constructing a regulatable version of the mouse p53 promoter that will be used to replace the endogenous promoter in mouse strain 129 by homologous recombination. Once crossed to lac repressor mice, we will be able to control the normal expression of p53.