PMID:2649886
| Citation |
Lovett, ST and Kolodner, RD (1989) Identification and purification of a single-stranded-DNA-specific exonuclease encoded by the recJ gene of Escherichia coli. Proc. Natl. Acad. Sci. U.S.A. 86:2627-31 |
|---|---|
| Abstract |
The Escherichia coli recJ gene product was overproduced using a plasmid that carries the recJ gene downstream of a strong regulatable promoter and a strong ribosome-binding site. Overexpression of recJ produced a concomitant increase in the levels of single-stranded-DNA-specific nuclease activity present in crude cell extracts. This nuclease activity was purified to homogeneity and found to reside in a 60-kDa polypeptide. This polypeptide was induced with recJ overexpression and had the size and N-terminal amino acid sequence identical to the predicted RecJ protein sequence. The RecJ nuclease degraded linear single-stranded DNA but did not have exonuclease activity on linear double-stranded substrates or endonuclease activity on either single-stranded or double-stranded substrates. The RecJ exonuclease had greater activity on duplex DNA molecules with 5'-rather than 3'-single-stranded tails. |
| Links | |
| Keywords |
Amino Acid Sequence; Base Sequence; DNA, Single-Stranded/metabolism; Escherichia coli/enzymology; Escherichia coli/genetics; Exodeoxyribonucleases/genetics; Exodeoxyribonucleases/isolation & purification; Gene Expression Regulation; Genes, Bacterial; Kinetics; Molecular Sequence Data; Mutation; Recombination, Genetic; Substrate Specificity |
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