Abstract
The chromosomal DNA molecules of a standard laboratory strain of Saccharomyces cerevisiae have been separated into 12 well-resolved bands by orthogonal-field-alternation gel electrophoresis. DNA X DNA hybridization probes derived from cloned genes have been used to correlate this banding pattern with yeast's genetically defined chromosomes. The 12 bands are shown to represent 9 singlets and 3 comigrating doublets, thereby accounting for 15 chromosomes that were identified as I-XI and XIII-XVI. Because the three comigrating doublets could be readily resolved in certain laboratory yeast strains that contain chromosome-length polymorphisms relative to our standard strain, all 15 of these chromosomes could be displayed as a single band in at least one of four strains that were studied. A 16th chromosome (number XII), which is known to contain the genes for rRNA, does not reproducibly enter the gels. By making use of the band identifications, the previously unmapped fragment F8 was assigned to chromosome XIII. With the possible exception of chromosomes that differ greatly in size or electrophoretic behavior from all the known chromosomes, the results appear to define a complete "electrophoretic karyotype" for yeast.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Beckmann J. S., Johnson P. F., Abelson J. Cloning of yeast transfer RNA genes in Escherichia coli. Science. 1977 Apr 8;196(4286):205–208. doi: 10.1126/science.322282. [DOI] [PubMed] [Google Scholar]
- Broach J. R. Construction of high copy yeast vectors using 2-microns circle sequences. Methods Enzymol. 1983;101:307–325. doi: 10.1016/0076-6879(83)01024-1. [DOI] [PubMed] [Google Scholar]
- Broach J. R., Friedman L., Sherman F. Correspondence of yeast UAA suppressors to cloned tRNASerUCA genes. J Mol Biol. 1981 Aug 15;150(3):375–387. doi: 10.1016/0022-2836(81)90553-2. [DOI] [PubMed] [Google Scholar]
- Broach J. R., Hicks J. B. Replication and recombination functions associated with the yeast plasmid, 2 mu circle. Cell. 1980 Sep;21(2):501–508. doi: 10.1016/0092-8674(80)90487-0. [DOI] [PubMed] [Google Scholar]
- Byers B., Goetsch L. Electron microscopic observations on the meiotic karyotype of diploid and tetraploid Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1975 Dec;72(12):5056–5060. doi: 10.1073/pnas.72.12.5056. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Carle G. F., Olson M. V. Separation of chromosomal DNA molecules from yeast by orthogonal-field-alternation gel electrophoresis. Nucleic Acids Res. 1984 Jul 25;12(14):5647–5664. doi: 10.1093/nar/12.14.5647. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Eibel H., Philippsen P. Preferential integration of yeast transposable element Ty into a promoter region. 1984 Jan 26-Feb 1Nature. 307(5949):386–388. doi: 10.1038/307386a0. [DOI] [PubMed] [Google Scholar]
- Fangman W. L. Separation of very large DNA molecules by gel electrophoresis. Nucleic Acids Res. 1978 Mar;5(3):653–665. doi: 10.1093/nar/5.3.653. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Goldring E. S., Grossman L. I., Krupnick D., Cryer D. R., Marmur J. The petite mutation in yeast. Loss of mitochondrial deoxyribonucleic acid during induction of petites with ethidium bromide. J Mol Biol. 1970 Sep 14;52(2):323–335. doi: 10.1016/0022-2836(70)90033-1. [DOI] [PubMed] [Google Scholar]
- Hartwell L. H. Macromolecule synthesis in temperature-sensitive mutants of yeast. J Bacteriol. 1967 May;93(5):1662–1670. doi: 10.1128/jb.93.5.1662-1670.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Horowitz H., Thorburn P., Haber J. E. Rearrangements of highly polymorphic regions near telomeres of Saccharomyces cerevisiae. Mol Cell Biol. 1984 Nov;4(11):2509–2517. doi: 10.1128/mcb.4.11.2509. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Langlois R. G., Yu L. C., Gray J. W., Carrano A. V. Quantitative karyotyping of human chromosomes by dual beam flow cytometry. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7876–7880. doi: 10.1073/pnas.79.24.7876. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Laughon A., Gesteland R. F. Primary structure of the Saccharomyces cerevisiae GAL4 gene. Mol Cell Biol. 1984 Feb;4(2):260–267. doi: 10.1128/mcb.4.2.260. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mortimer R. K., Hawthorne D. C. Genetic Mapping in Saccharomyces IV. Mapping of Temperature-Sensitive Genes and Use of Disomic Strains in Localizing Genes. Genetics. 1973 May;74(1):33–54. doi: 10.1093/genetics/74.1.33. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mortimer R. K., Schild D. Genetic map of Saccharomyces cerevisiae. Microbiol Rev. 1980 Dec;44(4):519–571. doi: 10.1128/mr.44.4.519-571.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Olson M. V., Loughney K., Hall B. D. Identification of the yeast DNA sequences that correspond to specific tyrosine-inserting nonsense suppressor loci. J Mol Biol. 1979 Aug 15;132(3):387–410. doi: 10.1016/0022-2836(79)90267-5. [DOI] [PubMed] [Google Scholar]
- Perkins D. D., Barry E. G. The cytogenetics of Neurospora. Adv Genet. 1977;19:133–285. doi: 10.1016/s0065-2660(08)60246-1. [DOI] [PubMed] [Google Scholar]
- Petes T. D. Yeast ribosomal DNA genes are located on chromosome XII. Proc Natl Acad Sci U S A. 1979 Jan;76(1):410–414. doi: 10.1073/pnas.76.1.410. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sandmeyer S. B., Olson M. V. Insertion of a repetitive element at the same position in the 5'-flanking regions of two dissimilar yeast tRNA genes. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7674–7678. doi: 10.1073/pnas.79.24.7674. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schwartz D. C., Cantor C. R. Separation of yeast chromosome-sized DNAs by pulsed field gradient gel electrophoresis. Cell. 1984 May;37(1):67–75. doi: 10.1016/0092-8674(84)90301-5. [DOI] [PubMed] [Google Scholar]
- Schwartz D. C., Saffran W., Welsh J., Haas R., Goldenberg M., Cantor C. R. New techniques for purifying large DNAs and studying their properties and packaging. Cold Spring Harb Symp Quant Biol. 1983;47(Pt 1):189–195. doi: 10.1101/sqb.1983.047.01.024. [DOI] [PubMed] [Google Scholar]
- Souciet J. L., Hubert J. C., Lacroute F. Cloning and restriction mapping of the yeast URA2 gene coding for the carbamyl phosphate synthetase aspartate-transcarbamylase complex. Mol Gen Genet. 1982;186(3):385–390. doi: 10.1007/BF00729458. [DOI] [PubMed] [Google Scholar]
- Tschumper G., Carbon J. Delta sequences and double symmetry in a yeast chromosomal replicator region. J Mol Biol. 1982 Apr 5;156(2):293–307. doi: 10.1016/0022-2836(82)90330-8. [DOI] [PubMed] [Google Scholar]
- Van der Ploeg L. H., Cornelissen A. W., Michels P. A., Borst P. Chromosome rearrangements in Trypanosoma brucei. Cell. 1984 Nov;39(1):213–221. doi: 10.1016/0092-8674(84)90207-1. [DOI] [PubMed] [Google Scholar]
- Van der Ploeg L. H., Schwartz D. C., Cantor C. R., Borst P. Antigenic variation in Trypanosoma brucei analyzed by electrophoretic separation of chromosome-sized DNA molecules. Cell. 1984 May;37(1):77–84. doi: 10.1016/0092-8674(84)90302-7. [DOI] [PubMed] [Google Scholar]
- Wickner R. B., Boutelet F., Hilger F. Evidence for a new chromosome in Saccharomyces cerevisiae. Mol Cell Biol. 1983 Mar;3(3):415–420. doi: 10.1128/mcb.3.3.415. [DOI] [PMC free article] [PubMed] [Google Scholar]