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Recombinant pseudorabies virus DNase exhibits a RecBCD-like catalytic function
C Y Hsiang1, T Y Ho, C H Hsiang
1Department of Microbiology, China Medical College, Taichung, Taiwan 40421, ROC.
The Biochemical Journal
|April 16, 1998
Summary
Pseudorabies virus (PRV) DNase was purified and characterized. This enzyme shows multiple nuclease activities, suggesting a potential role in PRV DNA recombination.
Area of Science:
- Molecular Biology
- Virology
- Enzymology
Background:
- The pseudorabies virus (PRV) DNase gene location is known.
- Further characterization of PRV DNase enzymatic properties is needed.
Purpose of the Study:
- To express and purify PRV DNase in Escherichia coli.
- To characterize the biochemical and enzymatic properties of PRV DNase in vitro.
Main Methods:
- PRV DNase gene was expressed in E. coli using a pET vector.
- The recombinant protein was purified to homogeneity.
- Nuclease activity assays were performed in vitro.
Main Results:
- Recombinant PRV DNase requires alkaline pH and Mg2+ ions.
- The enzyme exhibits endonuclease, 5'-exonuclease, and 3'-exonuclease activities on single- and double-stranded DNA.
- Activities are random with no significant base preference.
Conclusions:
- PRV DNase possesses multiple nuclease activities similar to enzymes involved in recombination.
- PRV DNase may play a role in recombination during PRV infection.