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On the DNA content and ploidy of trypanosomes
Authors:Piet Borst  Mels Van der Ploeg  Jos F.M. Van Hoek  Johan Tas  Jan James
Affiliation:1. Section for Medical Enzymology and Molecular Biology, Laboratory of Biochemistry, University of Amsterdam, Jan Swammerdam Institute, P.O. Box 60 000, 1005 GA Amsterdam, The Netherlands;2. Department of Histochemistry and Cytochemistry, State University of Leiden, Wassenaarseweg 72, 2333 AL Leiden, The Netherlands;3. Laboratory of Histology and Cell Biology, University of Amsterdam, Jan Swammerdam Institute, P.O. Box 60 000, 1005 GA Amsterdam, The Netherlands
Abstract:We have determined the nuclear and kinetoplast DNA content of two trypanosomatids by quantitative absorption and fluorescence cytophotometry of individual Feulgen-pararosaniline stained cells. For the insect trypanosomatid Crithidia fasciculata we find nuclear and kinetoplast DNA contents of 0.095 and 0.032 pg per non-replicating cell. For the African trypanosome Trypanosoma brucei these values are 0.097 and 0.004 pg. A sub-population of T. brucei cells with two kinetoplasts and one nucleus was found to contain 0.181 pg/nucleus. The DNA values of bloodstream form T. brucei and the procyclic culture form were not significantly different.In DNA-DNA renaturation experiments the haploid amount of DNA in T. brucei was previously found to be 0.041 pg/nucleus (Borst, P., Fase-Fowler, F., Frasch, A.C.C., Hoeijmakers, J.H.J. and Weijers, P.J. (1980) Mol. Biochem. Parasitol. 1, 221–246). Our data, therefore, indicate that T. brucei is diploid. No sub-population of haploid cells was observed in T. brucei grown in rats or in culture.
Keywords:Nuclear DNA content  Kinetoplast DNA content  Ploidy  Feulgen cytophotometry  kDNA  kinetoplast DNA  mtDNA  mitochondrial DNA  Hepes  4-(2-hydroxyethyl)-1-piperazineethanesulphonic acid
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