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Nucleic Acids Research, 1978, Vol. 5, No. 2 491-504
© 1978

Articles

Sequence arrangement of the 16S and 26S rRNA genes in the pathogenic haemoflagellate Leishmania donovani

Wilson Leon, David L. Fouts and Jerry Manning

Department of Molecular Biology and Biochemistry, University of California Irvine, CA 92717, USA

Received November 11, 1977. Kinetic and chemical analysis show that the haploid genome of Leishmania donovani has between 4.6 and 6.5 x 107 Kb pairs of DNA. Cot analysis shows that the genome contains 12% rapidly reassociating DNA, 13% middle repetitive DNA with an average reiteration frequency of 77 and 62% single copy DNA. Saturation hybridization experiments show that 0.82% of the nuclear DNA is occupied by rRNA coding sequences. The average repetition frequency of these sequences is determined to be 166. Sedimentation velocity studies indicate the two major rRNA species have sedimentation values of 26S and 16S, respectively. The arrangement of the rRNA genes and their spacer sequences on long strands of purified rDNA has been determined by the examination of the structure of rRNA:DNA hybrids prepared for electron microscopy by the gene 32-ethidium bromide technique. Long DNA strands are observed to contain several gene sets (16S + 26S). One repeat unit contains the following sequences in the order given: (a) A l6S gene of length 2.12 Kb, (b) An internal transcribed spacer (Spl) of length 1.23 Kb, which contains a short sequence that may code for a 5.8S rRNA (c) The 26S gene with a length of 4.31 Kb which contains an internal gap region of length 0.581 Kb, (d) An external spacer of average length 5.85 Kb.


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