| 摘要 |
PURPOSE:To obtain a DNA fragment containing catalase gene of fungus specifically induced in cultivation with n-alkane. CONSTITUTION:Total RNA is separated from a yeast belonging to the genus Candida, e.g. yeast cell Candida tropicalis grown in n-alkane and purified by applying oligo(dT)cellulose column chromatography to liberate poly(A)-containing RNA (mRNA). Corresponding cDNA library is obtained from the above- mentioned mRNA and the cDNA is methylated by DNA methylase to protect restriction enzyme cutting site and then DNA linker containing the above- mentioned restriction enzyme cutting site at the both end is added thereto and the added cDNA is digested by the restriction enzyme. The aimed cDNA is obtained by cloning the digested cDNA to cloning vector, infecting into Escherichia coli, cultivating in a culture media containing IPTG, etc. and screening formed plaque using an antibody which is specific to catalase of Candida yeast. Promoter of fungus catalase is useful in expressing an extraneous gene using a non-ethanol-fermentable carbon source. |
