PURPOSE:To massively prepare rat liver P450c25 by culturing Saccharomyces.cerevisiae AH22 strain holding a specific plasmid in a medium. CONSTITUTION:There is provided a novel expression plasmid pAC25 (14.1kb)(A) combined with a rat liver 450c25 gene at the downstream of the promoter of a yeast alcohol dehydrogenase gene and expressing the rat liver 450c25 in yeast. The component A and Saccharomyces.cerevisiae AH22 strain (B) are mixed and cultured to form a transformant, rat liver P450c25-producing strain(C), in which the component A has been introduced into the component B. The strain C is cultured in a medium containing glucose to prepare 5/1-cholestane3alpha, 7alpha, 12alpha, 27-tetraol and 1alpha, 25-(OH)2, vitamin D3.
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