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Carbon catabolite repression in yeast depends on catalytic active hexokinase isoenzyme PII ( Entian 1980a ). A yeast strain lacking hexokinase isoenzymes PI and PII was transformed, using a recombinant pool with inserts of yeast nuclear DNA up to 10 kbp in length. One hundred transformants for hexokinase were obtained. All selected plasmids coded for hexokinase isoenzyme PII, none for hexokinase isoenzyme PI, and carbon catabolite repression was restored in the transformants. Thirty-five independently isolated stable plasmids were investigated further. Analysis with the restriction enzyme EcoRI showed that these plasmids fell into two classes with different restriction behaviour. One representative of each class was amplified in Escherichia coli and transferred back into the yeast hexokinase-deficient strain with concomitant complementation of the nuclear mutation. The two types of insert were analysed in detail with 16 restriction enzymes, having 0-3 cleavage sites on transformant vector YRp7 . The plasmids differed from each other by the orientation of the yeast insert in the vector. After yeast transformation with fragments of one plasmid the hexokinase PII gene was localised within a region of 1.65 kbp.
K U Fröhlich, and
K D Entian, and
D Mecke
July 2001,
Yeast (Chichester, England),
K U Fröhlich, and
K D Entian, and
D Mecke
May 1989,
Journal of general microbiology,
K U Fröhlich, and
K D Entian, and
D Mecke
April 1998,
FEBS letters,
K U Fröhlich, and
K D Entian, and
D Mecke
September 1988,
Journal of general microbiology,
K U Fröhlich, and
K D Entian, and
D Mecke
December 1986,
Journal of general microbiology,
K U Fröhlich, and
K D Entian, and
D Mecke
March 1980,
Nucleic acids research,
K U Fröhlich, and
K D Entian, and
D Mecke
December 1981,
Gene,
K U Fröhlich, and
K D Entian, and
D Mecke
July 1997,
Biochemistry. Biokhimiia,
K U Fröhlich, and
K D Entian, and
D Mecke
January 1980,
Molekuliarnaia biologiia,
K U Fröhlich, and
K D Entian, and
D Mecke
June 1988,
Gene,
