Genetics, Vol 136, 849-856, Copyright © 1994

INVESTIGATIONS

Efficient Targeted Integration at leu1-32 and ura4-294 in Schizosaccharomyces pombe

J. B. Keeney and J. D. Boeke
Johns Hopkins University School of Medicine, Department of Molecular Biology and Genetics, Baltimore, Maryland 21205

Homologous integration into the fission yeast Schizosaccharomyces pombe has not been well characterized. In this study, we have examined integration of plasmids carrying the leu1(+) and ura4(+) genes into their chromosomal loci. Genomic DNA blot analysis demonstrated that the majority of transformants have one or more copies of the plasmid vector integrated via homologous recombination with a much smaller fraction of gene conversion to leu1(+) or ura4(+). Non-homologous recombination events were not observed for either gene. We describe the construction of generally useful leu1(+) and ura4(+) plasmids for targeted integration at the leu1-32 and ura4-294 loci of S. pombe.

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				L. D. Berry, A. Feoktistova, M. D. Wright, and K. L. Gould The Schizosaccharomyces pombe dim1+ Gene Interacts with the Anaphase-Promoting Complex or Cyclosome (APC/C) Component lid1+ and Is Required for APC/C Function Mol. Cell. Biol., April 1, 1999; 19(4): 2535 - 2546. [Abstract] [Full Text] [PDF]

				D. DeRyckere, C. L. Smith, and G. S. Martin The Role of Nucleotide Binding and Hydrolysis in the Function of the Fission Yeast cdc18+ Gene Product Genetics, April 1, 1999; 151(4): 1445 - 1457. [Abstract] [Full Text]

				V. D. Dang, M. J. Benedik, K. Ekwall, J. Choi, R. C. Allshire, and H. L. Levin A New Member of the Sin3 Family of Corepressors Is Essential for Cell Viability and Required for Retroelement Propagation in Fission Yeast Mol. Cell. Biol., March 1, 1999; 19(3): 2351 - 2365. [Abstract] [Full Text] [PDF]

				J. Bahler, A. B. Steever, S. Wheatley, Y.-l. Wang, J. R. Pringle, K. L. Gould, and D. McCollum Role of Polo Kinase and Mid1p in Determining the Site of Cell Division in Fission Yeast J. Cell Biol., December 14, 1998; 143(6): 1603 - 1616. [Abstract] [Full Text] [PDF]

				E. F. Hoff, H. L. Levin, and J. D. Boeke Schizosaccharomyces pombe Retrotransposon Tf2 Mobilizes Primarily through Homologous cDNA Recombination Mol. Cell. Biol., November 1, 1998; 18(11): 6839 - 6852. [Abstract] [Full Text]

				M. Penney, C. Wilkinson, M. Wallace, J.-P. Javerzat, K. Ferrell, M. Seeger, W. Dubiel, S. McKay, R. Allshire, and C. Gordon The pad1+ Gene Encodes a Subunit of the 26 S Proteasome in Fission Yeast J. Biol. Chem., September 11, 1998; 273(37): 23938 - 23945. [Abstract] [Full Text] [PDF]

				G. W. Brown and T. J. Kelly Purification of Hsk1, a Minichromosome Maintenance Protein Kinase from Fission Yeast J. Biol. Chem., August 21, 1998; 273(34): 22083 - 22090. [Abstract] [Full Text] [PDF]

				D. Bhaumik and T. S.-F. Wang Mutational Effect of Fission Yeast Polalpha on Cell Cycle Events Mol. Biol. Cell, August 1, 1998; 9(8): 2107 - 2123. [Abstract] [Full Text]

				M. K. Balasubramanian, D. McCollum, L. Chang, K. C. Y. Wong, N. I. Naqvi, X. He, S. Sazer, and K. L. Gould Isolation and Characterization of New Fission Yeast Cytokinesis Mutants Genetics, July 1, 1998; 149(3): 1265 - 1275. [Abstract] [Full Text] [PDF]

				D. A. Sherman, S. G. Pasion, and S. L. Forsburg Multiple Domains of Fission Yeast Cdc19p (MCM2) Are Required for Its Association with the Core MCM Complex Mol. Biol. Cell, July 1, 1998; 9(7): 1833 - 1845. [Abstract] [Full Text]

				J. Bähler and J. R. Pringle Pom1p, a fission yeast protein kinase that provides positional information for both polarized growth and cytokinesis Genes & Dev., May 1, 1998; 12(9): 1356 - 1370. [Abstract] [Full Text]

				A. Lopez-Girona, O. Mondesert, J. Leatherwood, and P. Russell Negative Regulation of Cdc18 DNA Replication Protein by Cdc2 Mol. Biol. Cell, January 1, 1998; 9(1): 63 - 73. [Abstract] [Full Text]

				L. D. Berry and K. L. Gould Fission Yeast dim1+ Encodes a Functionally Conserved Polypeptide Essential for Mitosis J. Cell Biol., June 16, 1997; 137(6): 1337 - 1354. [Abstract] [Full Text] [PDF]

				G. W. Brown, P. V. Jallepalli, B. J. Huneycutt, and T. J. Kelly Interaction of the S phase regulator Cdc18 with cyclin-dependent kinase in fission yeast PNAS, June 10, 1997; 94(12): 6142 - 6147. [Abstract] [Full Text] [PDF]