Establishment of Transgene Detection System with Mouse Tyrosinase Gene as a Visible Reporter Gene

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The transgene detection is the most time- and money- consuming step in transgenesis. Without this step, transgenesis must be proceeded more efficiently. The aim of our project is to examine the possibility whether tyrosinase minigene can be used to be a visible reporter gene in transgenesis. If succeeded, we can eliminate the transgene detection using genomic DNA extracted from tissue of transgenic candidate animals, which is essential but the most time- and money-consuming step in transgenesis. The conditions for the reporter gene are : 1) its stable expression in authentic tissues such as hair follicle in skin and eye, 2) minimum size as possible because that the reporter gene must be connected with other gene (s) of interest. For examine these respects, we have constructed a new tyrosinase minigene mg-Tyrs-JS with its truncated promoter region, 1.9 Kbp shortened than the previous minigene mg-Tyrs-J constructed by Tanaka et al. (Devel-opment 00 : 000-000,1990) . Introduced into cultured mouse L cells by electroporation method, the truncated minigene mg-Tyrs-Js was expressed in the cells. Then, we made transgenic mice using this minigene. Unexpectedly, however, no transgenic mice carrying with this minigene could be obtained with our great efforts, suggesting that the other tissue-specific regulatory element (s) must exist in the eliminated promoter region.