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Nucleic Acids Research, 1992, Vol. 20, No. 20 5351-5356
© 1992

MOLECULAR BIOLOGY

Characterization of Chlorella virus PBCV-1 CviAII restriction and modification system

Yanping Zhang, Michael Nelson, Joseph W. Nietfeldt, Dwight E. Burbank and James L. Van Etten*

Department of Plant Pathology, University of Nebraska Lincoln, NE 68583-0722, USA

* To whom correspondence should be addressed

Received July 20, 1992. Revised September 21, 1992. Accepted September 21, 1992.

A second DNA site-specific (restriction) endonuclease (R.CviAII) and its cognate adenine DNA methyltransferase (M.CviAII) were isolated from virus PBCV-1 infected Chlorella strain NC64A cells. R.CviAII, a heteroschizomer of the bacterial restriction endonuclease NlaIII, recognizes the sequence CATG, and does not cleave CmATG sequences. However, unlike NlaIII, which cleaves after the G and does not cleave either CmATG or mCATG sequences, CviAII cleaves between the C and A and is unaffected by mCATG methylation. The M.CviAII and R.CviAII genes were cloned and their DNA sequences were determined. These genes are tandemly arranged head-to-tail such that the TAA termination codon of the M.CviAII methyltransferase gene overlaps the ATG translational start site of R.CviAII endonuclease. R.CviAII is the first chlorella virus site-specific endonuclease gene to be cloned and sequenced.


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