Functional expression of the cre-lox site-specific recombination system in the yeast Saccharomyces cerevisiae.

The procaryotic cre-lox site-specific recombination system of coliphage P1 was shown to function in an efficient manner in a eucaryote, the yeast Saccharomyces cerevisiae. The cre gene, which codes for a site-specific recombinase, was placed under control of the yeast GALI promoter. lox sites flanking the LEU2 gene were integrated into two different chromosomes in both orientations. Excisive recombination at the lox sites (as measured by loss of the LEU2 gene) was promoted efficiently and accurately by the Cre protein and was dependent upon induction by galactose. These results demonstrate that a procaryotic recombinase can enter a eucaryotic nucleus and, moreover, that the ability of the Cre recombinase to perform precise recombination events on the chromosomes of S. cerevisiae is unimpaired by chromatin structure.

[1]  B. Sauer,et al.  Bacteriophage P1 cre gene and its regulatory region. Evidence for multiple promoters and for regulation by DNA methylation. , 1986, Journal of molecular biology.

[2]  R. Zagursky,et al.  Rapid and easy sequencing of large linear double-stranded DNA and supercoiled plasmid DNA , 1985 .

[3]  J. Rine,et al.  Regulated expression of endonuclease EcoRI in Saccharomyces cerevisiae: nuclear entry and biological consequences. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[4]  R. Hoess,et al.  Mechanism of strand cleavage and exchange in the Cre-lox site-specific recombination system. , 1985, Journal of molecular biology.

[5]  S. C. Falco,et al.  Genetic analysis of mutants of Saccharomyces cerevisiae resistant to the herbicide sulfometuron methyl. , 1985, Genetics.

[6]  R. Brent,et al.  A bacterial repressor protein or a yeast transcriptional terminator can block upstream activation of a yeast gene , 1984, Nature.

[7]  R. W. Davis,et al.  Sequences that regulate the divergent GAL1-GAL10 promoter in Saccharomyces cerevisiae , 1984, Molecular and cellular biology.

[8]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[9]  R. Peters.  Nuclear envelope permeability measured by fluorescence microphotolysis of single liver cell nuclei. , 1983, The Journal of biological chemistry.

[10]  R. Hoess,et al.  Studies on the properties of P1 site-specific recombination: Evidence for topologically unlinked products following recombination , 1983, Cell.

[11]  M. Kozak.  Comparison of initiation of protein synthesis in procaryotes, eucaryotes, and organelles. , 1983, Microbiological reviews.

[12]  D. Hanaham.  STUDIES ON TRANSFORMATION ON E. COLI WITH PLASMIDS , 1983 .

[13]  D. Hamer,et al.  Regulation of a metallothionein-growth hormone hybrid gene in bovine papilloma virus. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[14]  R. Rothstein.  One-step gene disruption in yeast. , 1983, Methods in enzymology.

[15]  I. Lehman,et al.  Immunological comparison of purified DNA polymerase alpha from embryos of Drosophila melanogaster with forms of the enzyme present in vivo. , 1982, The Journal of biological chemistry.

[16]  J. Broach.  The yeast plasmid 2μ circle , 1982, Cell.

[17]  N. Sternberg,et al.  Bacteriophage P1 site-specific recombination. I. Recombination between loxP sites. , 1981, Journal of molecular biology.

[18]  D. Holmes,et al.  A rapid boiling method for the preparation of bacterial plasmids. , 1981, Analytical biochemistry.

[19]  R. W. Davis,et al.  Rapid DNA isolations for enzymatic and hybridization analysis. , 1980, Methods in enzymology.

[20]  G R Stark,et al.  Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[21]  Ronald W. Davis,et al.  Isolation of galactose-inducible DNA sequences from Saccharomyces cerevisiae by differential plaque filter hybridization , 1979, Cell.

[22]  G. Fink,et al.  Methods in yeast genetics , 1979 .

[23]  G. Fink,et al.  Transformation of yeast. , 1978, Proceedings of the National Academy of Sciences of the United States of America.

[24]  F. Sanger,et al.  DNA sequencing with chain-terminating inhibitors. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[25]  P Berg,et al.  Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. , 1977, Journal of molecular biology.

[26]  M. Groudine,et al.  Chromosomal subunits in active genes have an altered conformation. , 1976, Science.

[27]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[28]  R. Axel,et al.  Analysis of DNA of isolated chromatin subunits. , 1975, Proceedings of the National Academy of Sciences of the United States of America.

[29]  B. G. Adams.  Induction of Galactokinase in Saccharomyces cerevisiae: Kinetics of Induction and Glucose Effects , 1972, Journal of bacteriology.

[30]  M Mandel,et al.  Calcium-dependent bacteriophage DNA infection. , 1970, Journal of molecular biology.

[31]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[32]  D. Hawthorne,et al.  ENZYMATIC EXPRESSION AND GENETIC LINKAGE OF GENES CONTROLLING GALACTOSE UTILIZATION IN SACCHAROMYCES. , 1964, Genetics.

[33]  8.7: Double-strand-break model for Recombination , 2022 .