Scientific papers // New enzymes
Restriction endonuclease Bis I from Bacillus subtilis T30 recognizes methylated sequence 5’-G(5mC)↓NGC-3’
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Translated from Biotechnologia (russ.). 2005. N 3. P. 22-26
Bacillus subtilis strain T30, producing a novel restriction endonuclease Bis I, has been isolated and characterized. The enzyme recognizes methylated DNA sequence 5’-G(5mC)↓NGC-3’ and cleaves it as it is shown by arrow. Due to cleavage of only modified DNAs Bis I may find a practical application in genetic engineering experiments as well as in determination of eukaryotic DNA methylation status.
PspXI, a novel restriction endonuclease that recognizes the unusual DNA sequence 5’-VC↓TCGAGB-3’
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Translated from Bulletin of biotechnology and physico-chemical biology named by Yu. A. Ovchinnikov V.1, 2005, No 1, pp 18-24
We have discovered a bacterial strain Pseudomonas species X11 that produces the unique restriction endonuclease PspXI. This enzyme recognizes the degenerate octanucleotide sequence 5’-VCTCGAGB-3’, where V stands for A, C or G and B stands for T, C or G. The PspXI restriction endonuclease preparation with the specific activity of 10000 units/ml was isolated using four chromatographic steps. PspXI cuts its recognition sequence between C and T producing cohesive ends compatible with those of produced by XhoI and SalI restriction endonucleases.
Restriction Endonuclease AjnI from Acinetobacter johnsonii R2, an Isoschizomer of EcoRII, recognizes 5'- CCWGG-3' and cleaves dcm-methylated sites
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Биотехнология, 2004, № 3, С. 19-24
A producer of restriction endonuclease AjnI has been isolated from natural resources and identified as bacterial species Acinetobacter johnsonii R2. The restriction enzyme purification and estimation of its activity is described. It has been shown that AjnI produces DNA fragments with 5'-CCWGG sticky ends similar to EcoRII, but cleavage is not blocked by dcm-methylation. A new restriction endonuclease AjnI may be widely used in genetic engineering.
A new site-specific DNA nickase N.Bst9I from Bacillus stearothermophilus 9
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Translated from BIOTEHNOLOGIA (Russia) 2001, No. 4, pp 3-8
Bacillus stearothermophilus 9 strain has been found during screening of bacteria from natural sources which produces a new site-specific nickase N.Bst9I. This enzyme recognizes and cleaves nucleotide sequence 5'-GAGTCNNNN^-3', so it is an isoschizomer of N.BstSEI endonuclease found earlier. But enzymatic properties of both enzymes are considerably different. N.Bst9I application in genetic engineering and biotechnology is more preferable because its "star" activity is much lower in comparison to N.BstSEI.
Read more: A new site-specific DNA nickase N.Bst9I from Bacillus stearothermophilus 9
BtrI, a novel restriction endonuclease, recognises the non-palindromic sequence 5'-CACGTC(-3/-3)-3'
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Nucleic Acids Research, 2000, Vol. 28, No. 11 E56-e56
The recognition sequence and cleavage positions of a new restriction endonuclease BtrI isolated from Bacillus stearothermophilus SE-U62 have been determined. BtrI belongs to a rare type IIQ of restriction endonucleases, which recognise non-palindromic nucleotide sequences and cleave DNA symmetrically within them. Type II restriction endonucleases (ENases) include a group of 53 prototypes that recognise non-palindromic DNA sequences (ENases with recognition sequences that are interrupted by more than one base pair are not considered). Cleavage positions have been determined for 45 of these [1]. Mainly, such ENases cleave DNA outside their recognition sites and are designated type IIS [2]. There are only a few so-called type IIQ restriction enzyme prototypes that cut both DNA strands symmetrically within their non-palindromic recognition sequences [3]. Here we report a new member of this rare subgroup of type II ENases.
PsiI, a unique Restriction Endonuclease Recognizing the DNA Sequence 5'-TTA^TAA-3'
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Translated from BIOCHEMISTRY (Russia) Vol. 64, No 4, pp. 481 - 482, 1999
PsiI, a new restriction endonuclease produced by the bacterial strain Pseudomonas sp. SE-G49, has been isolated and characterized. The enzyme cleaves DNA in the middle of its palindromic recognition sequence 5-TTA^TAA-3'. Thus, PsiI belongs to a rare group of type II restriction endonucleases whose recognition sites consist of AT base pairs only
AccBSI: A New Restriction Endonuclease from Acinetobacter calcoaceticus BS
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Translated from APPLIED BIOCHEMISTRY and MICROBIOLOGY (Russia) Vol. 33, No. 5, pp. 496-498, 1997
The recognition site of a new restriction endonuclease from Acinetobacter calcoaceticus BS was determined. This is a nonpalindromic sequence
3'-CTCGCC-5'
AccBSI restriction endonuclease cleaves DNA chains in the middle of the recognition sequence; therefore, ligation of its digestion fragments restores AccBSI recognition sites and generates palindromic sequences recognized by SacI and SacII restriction endonucleases.
Read more: AccBSI: A New Restriction Endonuclease from Acinetobacter calcoaceticus BS
N.BstSE, Site-Specific Nickase from Bacillus stearothermophilus SE-589
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Translated from “Molecular Biology”(Russia) 1996, Vol. 30, No. 6, pp. 1261-1267
A site-specific nickase recognizing and cleaving the DNA site
3'-C-T-C-A-G-N-N-N-N-N-N-5'
was isolated from Bacillus stearothermophilus SE-589 and named N.BstSE. Its properties indicate a possible relation with type II restriction endonucleases.
Read more: N.BstSE, Site-Specific Nickase from Bacillus stearothermophilus SE-589