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Nucleic Acids Research, 2003, Vol. 31, No. 14 e74
© 2003 Oxford University Press

A new Thermus sp. class-IIS enzyme sub-family: isolation of a ‘twin’ endonuclease TspDTI with a novel specificity 5'-ATGAA(N11/9)-3', related to TspGWI, TaqII and Tth111II

Piotr M. Skowron*,1,2, Jaroslaw Majewski1, Agnieszka Zylicz-Stachula1, Sylwia M. Rutkowska1, Izabela Jaworowska1 and Renata I. Harasimowicz-Slowinska1

1 EURx Ltd, Gdansk, Poland and 2 Department of Microbiology, Technical University of Gdansk, Gdansk, Poland

*To whom correspondence should be addressed at: EURx Ltd, ul. Jaskowa Dolina 29, 80–286 Gdansk, Poland. Tel: +48 58 345 2155; Fax: +48 58 341 7423; Email: piotrs{at}eurx.com.pl

The TspDTI restriction endonuclease, which shows a novel recognition specificity 5'-ATGAA(N11/9)-3', was isolated from Thermus sp. DT. TspDTI appears to be a ‘twin’ of restriction endonuclease TspGWI from Thermus sp. GW, as we have previously reported. TspGWI was isolated from the same location as TspDTI, it recognizes a related sequence 5'-ACGGA(N11/9)-3' and has conserved cleavage positions. Both enzymes resemble two other class-IIS endonucleases from Thermus sp.: TaqII and Tth111II. N-terminal amino acid sequences of TspGWI tryptic peptides exhibit 88.9–100% similarity to the TaqII sequence. All four enzymes were purified to homogeneity; their polypeptide sizes (114.5–122 kDa) make them the largest class-IIS restriction endonucleases known to date. The existence of a Thermus sp. sub-family of class-IIS restriction endonucleases of a common origin is herein proposed.


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