Takeda Chemical Industries, Inc.

Suntory Limited

Yasuji Oshima

Hiroyuki Araki

Toshihiko Ashikari

Keiko Sakakibara

Hiroto Kondo

Yoshinobu Kaneko

A DNA construct wherein a DNA fragment which is recombinable in yeast chromosomal DNA is directly or indirectly linked at both ends of a DNA fragment which comprises a pair of R sensitive sequences oriented in the same direction and flaking both an R gene placed under the control of an inducible promoter and an expressible selective marker gene, which is a DNA construct designed with the R sensitive sequences non-symmetrically shortened, so that no functionable R sensitive sequence remains after the R sensitive sequence recombination has occurred by expression of the R gene and the selective marker has been removed. Since no functionable R sensitive sequence remains after removal of the selective marker, recombination does not occur again, and thus the same selective marker may be used for multiple insertions of foreign genes.

Method for constructing transformant not having selective marker gene

The present invention relates to a DNA fragment having a promoter activity of PHO81 gene regulating the production of phosphatase, and which is obtainable from Saccharomyces cerevisiae.; a DNA fragment bearing a structural gene downstream from the above PHO81 promoter; a transformant containing a DNA fragment bearing the above PHO81 promoter; a transformant containing a DNA fragment bearing the above PHO81 promoter and a structural gene downstream from the PHO81 promoter; and a process for obtaining a gene product which includes culturing a transformant containing a DNA fragment bearing the above PHO81 promoter and a structural gene located downstream therefrom in a suitable medium until the gene product is formed and recovering the gene product from the culture. Pharmacologically important proteinous materials may be efficiently produced with the use of the above-described novel and potent promoter obtained from yeast which is a eukaryotic microorganism.

PhO81 promotor of Saccharomyces cerevisiae and use thereof for

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Takatsuki, Japan

Yasuji Oshima