The restriction endonuclease PecI from the bacterial strain Paracoccus species 12 recognizes the DNA sequence 5’-TTA^TAA-3’ and is an isoschizomer of PsiI

D.A. Gonchar¹, V.A. Chernukhin¹, V.S. Dedkov¹, N.A. Mikhnenkova¹, M.A. Abdurashitov¹, O.A. Belichenko¹, S.Kh. Degtyarev¹

¹ SibEnzyme Ltd, Novosibirsk

Corresponding author: Danila Gonchar, SibEnzyme Ltd, 2/12 Timakova St., Novosibirsk, 630117, Russia; Tel.: +7(383)333-4991; Fax: +7(383)333-6853; E-mail: gonchar@sibenzyme.ru

We identified a bacterial strain, Paracoccus species 12, which produces a novel restriction endonuclease named PecI. This enzyme recognizes the hexanucleotide palindromic DNA sequence 5’-TTA^TAA-3’ and cleaves it, as indicated by the arrow, generating blunt ends. Thus, PecI is a true isoschizomer of the restriction enzyme PsiI [1].

A preparation of PecI restriction endonuclease with a concentration of 10,000 U/mL was obtained through a four-step chromatographic purification process. The optimal reaction conditions for the enzyme are SE-buffer 5 (Y) (33 mM Tris-acetate (pH 7.9), 10 mM Mg-acetate, 66 mM K-acetate, 1 mM DTT) and a temperature of 37°C.

Used abbreviations:
BSA – bovine serum albumin; DTT – dithiothreitol; PAGE – polyacrylamide gel electrophoresis; bp – base pairs; RM system – restriction-modification system; Tris – tris(hydroxymethyl)aminomethane; EDTA – ethylenediaminetetraacetic acid; RE – restriction endonuclease.

Keywords: restriction endonucleases, isoschizomers

DOI:

Citation details:
D.A. Gonchar, V.A. Chernukhin, V.S. Dedkov, N.A. Mikhnenkova, M.A. Abdurashitov, O.A. Belichenko, S.Kh. Degtyarev. (2024) The restriction endonuclease PecI from the bacterial strain Paracoccus species 12 recognizes the DNA sequence 5’-TTA^TAA-3’ and is an isoschizomer of PsiI, DNA processing enzymes, Volume 2024(1), DOI:

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