Abstract
Bacterial plasmids containing no detectable homology with yeast DNA sequences were inserted into the yeast genome by cotransforming with a plasmid containing a yeast gene. Analysis of the yeast transformants confirmed that recombination events occurred between the prokaryotic sequences shared by the two plasmids and between the yeast sequences common to the cotransforming plasmid and to the genome. Multiple copies of the two plasmids, in both tandem and interspersed arrays, are inserted by this method. Populations of cells grown from individual transformants are heterogenous for the number of integrated sequences. The number of integrated bacterial sequences is greatly reduced after 100 generations of growth in the populations that initially contained large numbers of sequences, while it is stable in those populations that initially contained either a single or a small number of copies.
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References
Beach D, Piper M, Shall S (1980) Isolation of chromosomal origins of replication in yeast. Nature 284:185–187
Broach JR, Strathein JN, Hicks JB (1979) Transformation in yeast: development of a hybrid cloning vector and isolation of the CanI gene. Gene 8:121–133
Chan CSM, Tye B-K (1980) Autonomously replicating sequences in Saccharomyces cerevisiae. Proc Natl Acad Sci USA 77:6329–6333
Denhardt DT (1966) A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Comm 23:641–646
Hicks JB, Hinnen A, Fink GR (1978) Properties of yeast transformation. Cold Spring Harbor Symposia 43:1305–1313
Hinnen A, Hicks JB, Fink GR (1978) Transformation of yeast. Proc Natl Acad Sci USA 75:1929–1933
Jaenisch R, Jahner D, Nobis P, Simon I, Lohler J, Harbers K, Grotkopp D (1981) Chromosomal position and activation of retrovial genomes inserted into the germ line of mice. Cell 24:519–529
Kafatos FC, Jones CW, Efstratiadis A (1979) Determination of nucleic acid sequence homologies and relative concentrations by a dot hybridization procedure. Nucleic Acids Res 7:1541–1552
Kretschmer PJ, Chang ACY, Cohen SN (1975) Indirect selection of bacterial plasmids lacking identifiable properties. J Bacteriol 124:225–231
Rigby PWJ, Dieckmann M, Rhodes C, Berg P (1977) Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113:237–251
Sherman F, Fink G, Hicks J (1979) Laboratory manual for methods in yeast genetics. Cold Spring, Harbor Laboratory, Cold Spring Harbor, NY
Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517
Wigler M, Sweet R, Sim GK, Wold B, Pellicer A, Lacy E, Maniatis T, Silverstein S, Axel R (1979) Transformation of mammalian cells with genes from procaryotes and eucaryotes. Cell 16:777–785
Zamir A, Maina CV, Fink GR, Szalay AA (1981) Stable chromosomal integration of the entire nitrogen fixation gene cluster from Klebsiella pneumoniae in yeast. Proc Natl Acad Sci USA 78:3496–3500
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Communicated by H. Böhme
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Compton, J.L., Zamir, A. & Szalay, A.A. Insertion of nonhomologous DNA into the yeast genome mediated by homologous recombination with a cotransforming plasmid. Molec Gen Genet 188, 44–50 (1982). https://doi.org/10.1007/BF00332994
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DOI: https://doi.org/10.1007/BF00332994