Park S M, Ohkuma M, Masuda Y, Ohta A, Takagi M
Department of Biotechnology, University of Tokyo, Japan.
Yeast. 1997 Jan;13(1):21-9. doi: 10.1002/(SICI)1097-0061(199701)13:1<21::AID-YEA58>3.0.CO;2-L.
The GAL1 and GAL10 gene cluster encoding the enzymes of galactose utilization was isolated from an asporogenic yeast, Candida maltosa. The structure of the gene cluster in which both genes were divergently transcribed from the central promoter region resembled those of some other yeasts. The expression of both genes was strongly induced by galactose and repressed by glucose in the medium. Galactose-inducible expression vectors in C. maltosa were constructed on low- and high-copy number plasmids using the promoter regions of both genes. With these vectors and the beta-galactosidase gene from Kluyveromyces lactis as a reporter, galactose-inducible expression was confirmed. Homologous overexpression of members of the cytochrome P-450 gene family in C. maltosa was also successful by using a high-copy-number vector under the control of these promoters.
从产孢酵母麦芽糖假丝酵母中分离出了编码半乳糖利用酶的GAL1和GAL10基因簇。该基因簇中两个基因从中央启动子区域反向转录,其结构与其他一些酵母的相似。两个基因的表达在培养基中受到半乳糖的强烈诱导和葡萄糖的抑制。利用这两个基因的启动子区域,在低拷贝数和高拷贝数质粒上构建了麦芽糖假丝酵母中的半乳糖诱导表达载体。使用这些载体以及乳酸克鲁维酵母的β-半乳糖苷酶基因作为报告基因,证实了半乳糖诱导表达。通过使用在这些启动子控制下的高拷贝数载体,麦芽糖假丝酵母中细胞色素P-450基因家族成员的同源过表达也取得了成功。