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Nucleic Acids Research, 1989, Vol. 17, No. 8 3217-3227
© 1989

MOLECULAR BIOLOGY

Improved separation of chromosome-sized DNA from Trypanosoma brucei, stock 427-60

Lex H.T. Van der Ploeg, Cassandra L. Smith1, Ramona I. Polvere and Keith M. Gottesdiener

Department of Genetics and Development, College of Physicians and Surgeons, Columbia University 701 West 168 St., New York, NY 10032, USA 1Department of Microbiology and Psychiatry, College of Physicians and Surgeons, Columbia University 701 West 168 St., New York, NY 10032, USA

Received November 16, 1988. Revised March 8, 1989. Accepted March 8, 1989.

Separation of chromosome-sized DNA from the parasitic protozoan Trypanosoma brucei had previously resulted in the fractionation of DNA molecules that ranged in size from 50 kb up to roughly 1.5 Mb. The number of larger chromosomes and their size, accounting for 80% of the DNA of T.brucei remained unclear. We have now size separated these larger DNA molecules by pulsed field gel electrophoresis (PFG) and resolve a total of 20 bands, accounting for roughly 120 chromosomes, ranging in size from 50 kb up to the size of the largest, 5.7 Mb chromosome of Schizosaccharomyces pombe Three different VSG gene expression sites were located to chromosomes of 430 kb, 1.5 Mb and 3 Mb, respectively. We have not been able to identify additional, previously cryptic DNA rearrangements, that could explain the activation or inactivation of the expression sites.


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