Characterization of kinetoplast DNA networks from the insect trypanosome Crithidia luciliae

P. O. Weislogel, J. H.J. Hoeijmakers, A. H. Fairlamb, C. M. Kleisen, P. Borst

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20 Citaten (Scopus)


1. We have used the restriction endonucleases EcoRI and PstI to further characterize the structural components of intact kinetoplast DNA networks from stationary phase Crithidia luciliae. 2. Endonuclease PstI cuts less than 7% of the mini-circles (the major component in the network) and appears to give a single cut in the maxi-circles, allowing the isolation of this minor component in linearized form. The molecular weight of these linearized maxi-circles, determined by electron microscopy with phage PM2 DNA as internal standard, is 22 · 106. 3. Electron micrographs of intact networks show DNA considerably longer than the mini-circle contour length (0.8 μm) either in the network or attached to the edge. This long DNA never exceeds the size of maxi-circles (10.2 μm) and it is completely removed by treatment with either endonuclease PstI or endonuclease EcoRI (which cuts the maxi-circles and about 20% of the mini-circles). We conclude that the long DNA in these networks represent maxi-circles and that long circular oligomers of mini-circles are (virtually) absent. 4. Complete removal of maxi-circles with endonucleases PstI or EcoRI has little effect on the overall structure of the network. This shows that the mini-circles are not held together on maxi-circle strings.

Originele taal-2Engels
Pagina's (van-tot)167-179
Aantal pagina's13
TijdschriftBBA Section Nucleic Acids And Protein Synthesis
Nummer van het tijdschrift2
StatusGepubliceerd - 20 sep. 1977
Extern gepubliceerdJa


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