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http://dx.doi.org/10.5423/PPJ.2009.25.1.047

Molecular Characterization of a Nuclease Gene of Chlorella Virus SS-2  

Park, Yun-Jung (Department of Microbiology, Pukyong National University)
Jung, Sang-Eun (Department of Microbiology, Pukyong National University)
Choi, Tae-Jin (Department of Microbiology, Pukyong National University)
Publication Information
The Plant Pathology Journal / v.25, no.1, 2009 , pp. 47-53 More about this Journal
Abstract
Sequence analysis of the Chlorella virus SS-2 revealed one putative nuclease gene that is 807 bp long and encodes a 31kDa protein. Multiple sequence alignment analysis reveals the presence of highly conserved PD-(D/E)XK residues in the encoded protein. The gene cloned into an expression vector was expressed as a His-tagged fusion protein in chaperone containing pKJE7 cells. The recombinant protein was purified using a His-Trap chelating HP column and used for functional analysis. Exonuclease activity of the SS-2 nuclease was detected when the DNA substrates, such as linear ssDNA, PCR amplicon, linear dsDNA with 5'-overhang ends, 3'-overhang ends, or blunt ends were used. Covalently closed circular DNA was also degraded by the SS-2 recombinant protein, suggesting that the SS-2 nuclease has an endonuclease activity. Stable activity of SS-2 nuclease was observed between $10^{\circ}C$ and $50^{\circ}C$. The optimum pH concentrations for the SS-2 nuclease were pH 6.0-8.5. Divalent ions inhibited the SS-2 nuclease activity.
Keywords
algal virus; Chlorella virus; nuclease gene; recombinant protein;
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