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Pakistan Journal of Biological Sciences
  Year: 1999 | Volume: 2 | Issue: 4 | Page No.: 1299-1304
DOI: 10.3923/pjbs.1999.1299.1304
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Purification and Characterization of New Type II Restriction Endonucleases

Shahnaz Choudhry

Twelve bacterial strains were screened for the presence of new type II restriction endonucleases. Two are reported positive for the enzyme activity, and designated as BthDI, SarI, from Bacillus thuringiensis D4 (CAMB 2657) and Streptomyces aureomonopodiales (ATCC 19825). These enzymes have been purified by a combination of gel filtration, ion exchange and affinity chromatography. The DNA sequences recognized by these enzymes have been determined by analysis of the fragment patterns generated on different substrate DNAs, Lambda, T7, pUC19, pBR322, PhiX174RFI, and Adenovirus-2 (Ad-2). The cleavage sites within the recognition sequences have also been established by primed synthesis. Accordingly, the newly discovered enzymes BthDI and SarI recognize and cleave 4-6 nucleotide long stretch of DNA, CC!AGG, AGG! CCT, respectively.
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How to cite this article:

Shahnaz Choudhry , 1999. Purification and Characterization of New Type II Restriction Endonucleases . Pakistan Journal of Biological Sciences, 2: 1299-1304.

DOI: 10.3923/pjbs.1999.1299.1304






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