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Nucleic Acids Research, 1989, Vol. 17, No. 7 2801-2818
© 1989

MOLECULAR BIOLOGY

Construction of a Not I restriction map of the fission yeast Schizosaccharomyces pombe genome

Jian-Bing Fan, Yuji Chikashige2, Cassandra L. Smith1, Osami Niwa2, Mitsuhiro Yanagida2 and Charles R. Cantor

Department of Genetics and Development, College of Physicians and Surgeons, Columbia University 701 West 168th Street, New York, NY 10032, USA 1Department of Microbiology and Psychiatry, College of Physicians and Surgeons, Columbia University 701 West 168th Street, New York, NY 10032, USA 2Department of Biophysics, Faculty of Science, Kyoto University Sakyo-ku, Kyoto 606, Japan

Received November 4, 1988. Revised February 24, 1989. Accepted February 24, 1989.

Pulsed field gel electrophoresis and large DNA technology were used to construct a Not I restriction map of the entire genome of the fission yeast Schizosaccharomyces pombe. There are 14 detectable Not I sites in S. pombe 972h: 9 sites on chromosome I and 5 sites on chromosome II, while no Not I sites were found on chromosome III. The 17 fragments (including intact chromosome III) generated by Not I digestion were resolved by PFG electrophoresis. These fragments ranged in size from 4.5 kb to approximately 3.5 Mb. Various strategies were applied in determining, efficiently, the order of the fragments on the chromosomes. The genomic size measured by adding all the fragments together is about 14 Mb and the sizes of the three chromosomes are I, 5.7 Mb, II, 4.6 to 4.7 Mb, and III, 3.5 Mb. These are generally somewhat smaller than estimated previously.


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