Abstract
ABSTRACT The kinetoplast DNA (kDNA) of the hemoflagellate protozoa consists of a single large network of thousands of catenated minicircles and a smaller number of catenated larger maxicircles. In this report we briefly review the known information on the replication of the kDNA and present some recent results on characterization of the maxicircle component of the kDNA. Maxicircle DNA has been isolated by release from the network by cleavage with a restriction enzyme and separation on a buoyant density basis in CsCl in the presence of the dye, Hoechst 33258. In general the maxicircle DNA is higher in %A+T than the minicircle DNA. A restriction map of the maxicircle of Leishmania tarentolae has been constructed and the 9 and 12S kinetoplast RNA genes localized on this map. Several maxicircle fragments have been cloned in the bacterial plasmid, pBR322, including a 4.4 × 10 dalton Eco RI/BamHI fragment that contains the 9 and 12S RNA genes. Unit length minicircles have also been cloned in pBR322. The restriction maps of two different semi-homologous minicircle classes have been obtained. Similar studies have been performed on the kDNA of the related species, Trypanosoma brucei, which undergoes a cyclical regression and biogenesis of the mitochondrion during its life cycle.
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