| 摘要 |
PROBLEM TO BE SOLVED: To provide a method for economically producing an active alkaline phosphatase derived from a eukaryote by a large expression amount. SOLUTION: This method for producing a eukaryotic alkaline phosphatase in a yeast includes the steps of (a) cloning one gene sequence into different vectors, (b) transforming a yeast, (c) expression, and (d) purifying the alkaline phosphatase, and is characterized by amplifying, by the multiple transformation, the gene copy number of a transformant that a resistance genome to the first selection marker and the above gene sequence are integrated into the genome, adding the second vector having a resistance gene to the second selection marker, increasing the gene copy number by the multiple transformation by the second vector, wherein the multiple transformant is selected by the growth in a nutrient broth having an increased selection pressure, and selecting a clone in which copies of the above both selection markers-resistant gene and the above gene sequence are stably integrated. |
