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(Received for publication, September 20, 1995, and in revised form, June 3, 1996)
,
and¶
From the DNA end joining is a type of illegitimate
recombination characterized by the joining of two DNA ends that lack
homology. Using oligonucleotides as substrate, we found that an
exonuclease-free derivative of the Klenow fragment of
Escherichia coli DNA polymerase I can mediate DNA end
joining in vitro. DNA sequence analysis of product DNA
indicated that overlap products were formed between direct repeat
sequences at the termini of the oligonucleotides. Formation of
recombinant products was dependent on the strandedness of the substrate
DNA, and the rate of product formation was dependent on the size of the
potential overlap. With one to three complementary bases available for
pairing at the 3
Laboratory of Radiobiology and Environmental
Health and the ¶ Department of Radiation Oncology, University
of California, San Francisco, California 94143
termini, there was an absolute requirement that one
of the oligonucleotides be double-stranded, whereas with four
complementary bases, products were also formed in reactions with
single-stranded oligonucleotides. When noncomplementary nucleotides
were added to the terminus of one of the oligonucleotides, product
formation was delayed but not blocked. These data indicate that a DNA
polymerase can mediate DNA double strand break rejoining in the absence
of other proteins.
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