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PubMed1995

Analysis of recombination junctions in extrachromosomal circular DNA obtained by in-gel competitive reassociation.

Essentially all eukaryotic cells contain circular extrachromosomal DNA as a result of excision from the chromosomes. To obtain insight into the nature of recombination associated with the occurrence of such DNA species and its biological significance, we analyzed a library enriched in recombination junctions which was constructed by a novel DNA subtraction technique; in-gel competitive reassociation (IGCR). Furthermore, we also introduced inverse PCR to characterize chromosomal DNA fragments containing the recombination junctions. At least 45% of the clones in the library constructed by the IGCR procedure comprised DNA with recombination junctions. Nucleotide sequence analysis of the recombination junctions indicated that three of four extrachromosomal DNAs thus analyzed were produced through recombination between sequences with a 3-5 bp homology in the chromosomes. One extrachromosomal DNA was apparently generated through non-homologous recombination, possibly by end-to-end joining. These results have demonstrated the usefulness of IGCR in concentrating recombination junctions, which provide the most direct evidence for the mechanism of the recombinational events involved, from highly complex genomes.

Base Sequence↗
PubMed1993

Identification of a Babesia bovis gene with homology to the small subunit ribosomal RNA gene from the 35-kilobase circular DNA of Plasmodium falciparum.

Isolation of a 552-base pair (bp) fragment of a putative extrachromosomal small subunit ribosomal RNA (SSrRNA) gene from Babesia bovis was achieved using the polymerase chain reaction (PCR) followed by cloning and sequencing of the PCR product. The sequences of the oligonucleotide primers used for the PCR were derived from selected known sequences in the organellar SSrRNA gene which is encoded within the 35-kilobase (kb) circular DNA from Plasmodium falciparum. Comparison of the sequence of the 552-bp fragment from B. bovis with gene sequences from other organisms showed 71% identity with the organellar SSrRNA gene from P. falciparum and up to 65% identity with the plastid SSrRNA gene sequences from various other organisms. We conclude that the 552-bp fragment amplified by PCR from B. bovis is possibly derived from an organellar genome of this parasite.

Animals↗