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
aw Majewski1
ylicz-Stachula1
owi
ska1
1 EURx Ltd, Gda
sk, Poland and
2 Department of Microbiology, Technical University of Gda
sk, Gda
sk, Poland
*To whom correspondence should be addressed at: EURx Ltd, ul. Ja
kowa Dolina 29, 80286 Gda
sk, 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 88.9100% similarity to the TaqII sequence. All four enzymes were purified to homogeneity; their polypeptide sizes (114.5122 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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