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New DNA methyltransferase M.AgsI produces TTSA(m6A)

V.S.Dedkov, D.A.Gonchar, V.A.Chernukhin, M.A.Abdurashitov, S.G.Udalyeva, L.A.Urumceva, S.Kh.Degtyarev 
BioTechnology : An Indian Journal Volume 12 Issue 2, 2016 Pages: 59-112

Agrococcus species 25 DNA was cloned in pUC19 plasmid of Escherichia coli. Cloned DNA fragment contained two Opened Reading Frames with 8 amino acid motives which belonged to amino DNA methyltransferases. Thus M.AgsI can be the first of subunit adenine-(N6)-DNA methyltransferase. The enzyme was purified from the recombinant strain by chromatography on P-11 Phosphocellulose, Heparin-Sepharose and Hydroxyapatite. M.AgsI specificity was determined by a study of protection of lambda DNA methylated with M.AgsI against cleavage with some restriction endonucleases. A sensitivity of restriction endonucleases to M.AgsI-methylation was studied.

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A new methyl-directed site-specific DNA endonuclease MteI cleaves nonanucleotide sequence 5’-G(5mC)G(5mC)^NG(5mC)GC-3’/3’-CG(5mC)GN^(5mC)G(5mC)G-5’

This email address is being protected from spambots. You need JavaScript enabled to view it. , D.A. Gonchar, E.V. Kileva, V.A. Sokolova, L.N. Golikova, V.S. Dedkov, N.A. Mikhnenkova, S.Kh. Degtyarev

Translated from "Ovchinnikov bulletin of biotechnology and physical and chemical biology" V.8, No 1, pp 16-26, 2012

 

We have discovered and purified a new methyl-directed site-specific DNA endonuclease MteI from bacterial strain Microbacterium testaceum 17B. The enzyme recognizes methylated DNA sequence and doesn’t cleave unmethylated DNA. MteI is a first methyl-directed site-specific DNA endonuclease recognizing a prolonged DNA sequence and its activity depends on a number of 5-methycytosines and their positions in the recognition site. MteI cleaves DNA sequence 5’-G(5mC)G(5mC)↑NG(5mC)GC-3’/3’-CG(5mC)GN↓(5mC)G(5mC)G-5’ as indicated by arrows and this nonanucleotide is a minimal recognition site. The enzyme activity is significantly higher if 5’-GC-3’ dinucleotides in this site are replaced by 5’-G(5mC)-3’ dinucleotides and additional 5’-G(5mC)-3’ dinucleotides are present at 5’-ends in both DNA strands. Due to an ability to cleave only prolonged methylated DNA sequences MteI may find a practical application in the molecular biology and epigenetics studies. 

Read more: A new methyl-directed site-specific DNA endonuclease MteI cleaves nonanucleotide sequence...

A new site-specific methyl-directed DNA endonuclease PkrI recognizes and cuts methylated DNA sequence 5'-GCN^GC-3'/3'-CG^NCG-5' carrying at least three 5-methylcytosines

This email address is being protected from spambots. You need JavaScript enabled to view it. , T. N. Nayakshina, D.A. Gonchar, J.E. Tomilova, M.V. Tarasova, V.S. Dedkov, N.A. Mikhnenkova, S.Kh. Degtyarev

Translated from "Ovchinnikov bulletin of biotechnology and physical and chemical biology" V.7, No 3, pp 35-42, 2011

 

We have discovered and purified a new methyl-directed site-specific DNA endonuclease PkrI from bacterial strain Planomicrobium koreense 78k. PkrI recognizes and cuts methylated DNA sequence 5'-GCNGC-3'/3'-CGNCG-5' carrying at least three 5-methylcytosimes and doesn’t cleave unmethylated DNA. Due to its ability to cleave only modified DNA PkrI may find a practical application in genetic engineering experiments as well as in determination of DNA methylation status.

Read more: A new site-specific methyl-directed DNA endonuclease PkrI recognizes and cuts methylated DNA...

A new methyl-directed site-specific endonuclease KroI recognizes and cuts DNA sequence 5’-G^C(5mC)GGC-3’

This email address is being protected from spambots. You need JavaScript enabled to view it. , E.V. Kileva, J.E. Tomilova, A.A. Boltengagen, V.S. Dedkov, N.A. Mikhnenkova, D.A. Gonchar, L.N. Golikova, S.Kh. Degtyarev

Translated from "Ovchinnikov bulletin of biotechnology and physical and chemical biology" V.7, No 1, pp 14-20, 2011

 

We have discovered and purified a new methyl-directed site-specific DNA endonuclease KroI from bacterial strain Kocuria rosea 307. KroI recognizes and cuts DNA sequence 5’-G↑C(5mC)GGC-3’/3’-CGG(5mC)C↓G-5’ and doesn’t cleave unmethylated DNA. A new enzyme cleaves both strands of the recognition sequence if one or two central cytosines are methylated. KroI is a first methyl-directed site-specific DNA endonuclease that recognizes the non-degenerate hexanucleotide sequence. Due to its ability to cleave only modified DNA KroI may find a practical application in genetic engineering experiments as well as in determination of DNA methylation status.

Read more: A new methyl-directed site-specific endonuclease KroI recognizes and cuts DNA sequence...

Substrate specificity of new restriction endonuclease SetI

This email address is being protected from spambots. You need JavaScript enabled to view it. , Degtyarev S.Kh

 

 

A few site-specific endonucleases are currently known which recognize and cleave short (3- and 4-letter) degenerate DNA sequence. These are restriction endonucleases CviJI and different isoshizomers from eukaryotic Chlorella virus, which recognizes and cleaves the nucleotide sequence 5'-RGCY-3' [1]. In the present work we describe the substrate specificity of a new site specific endonuclease SetI, which is capable to cleave DNA sequence 5'- ASST^ -3' (four main sequences: ACGT^, AGCT^, ACCT^ and AGGT^). SetI was isolated from an E.coli strain that carries the cloned SetI gene from Streptomyces werraensis 37

 

Read more: Substrate specificity of new restriction endonuclease SetI

Substrate specificity of new restriction endonuclease FaiI

This email address is being protected from spambots. You need JavaScript enabled to view it. , Chernukhin V.A., Tomilova J.E., Degtyarev S.Kh

 

 

A few site-specific endonucleases are currently known which recognize and cleave short (3- and 4-letter) degenerate DNA sequence. These are restriction endonucleases CviJI and different isoshizomers from eukaryotic Chlorella virus, which recognizes and cleaves the nucleotide sequence 5'-RGCY-3' [1]. In the present work we describe the substrate specificity of a new site specific endonuclease FaiI, which is capable to cleave degenerate 5'-YATR-3' (four main sequences: CA^TG, CA^TA, TA^TG and TA^TA) and with the less effectiveness the majority other variants of three-letter sequence 5'-YAT-3' . This enzyme was isolated from bacterial strain Flavobacterium aquatile by the chromatographic methods

 

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New Restriction Endonuclease AluBI from Arthrobacter luteus B - AluI Isoshizomer, nonsensitive to Presence of 5-methylcytosine in the Recognition Sequence AGCT

This email address is being protected from spambots. You need JavaScript enabled to view it. , A. A. Boltengagen, G. V. Tarasova, V.S. Dedkov, S.Kh. Degtyarev

Translated from "Ovchinnikov bulletin of biotechnology and physical and chemical biology" V.3, No 1, pp 21-27, 2007

 

A new restriction endonuclease AluBI has been discovered and characterized. AluBI is an isoshizomer of well known restriction endonuclease AluI, which recognizes DNA sequence AGCT. Unlike AluI, enzyme AluBI is able to cleave DNA when recognition sequence contains 5-methylcytosines. AluBI also cleaves recognition sequence with two methylated cytosines, one modified at N4 and other at C5 positions. However, new enzyme doesn’t cleave DNA with two N4-methylcytosines in the recognition site.

Читать полный текст статьи New Restriction Endonuclease AluBI from Arthrobacter luteus B - AluI Isoshizomer, nonsensitive to...

Site-specific endonuclease BlsI recognizes DNA sequence 5’-G(5mC)N^GC-3’ and cleaves it producing 3’ sticky ends

This email address is being protected from spambots. You need JavaScript enabled to view it. , Yu.E. Tomilova, E.V. Chmuzh, O.O. Sokolova, V.S. Dedkov, S.Kh. Degtyarev

Translated from "Ovchinnikov bulletin of biotechnology and physical and chemical biology" V.3, No 1, pp 28-33, 2007

 

A bacterial strain Bacillus simplex 23, a producer of a site-specific endonuclease BlsI has been discovered. BlsI recognizes the methylated DNA sequence 5’-G(5mC)N↓GC-3’, like the earlier described site-specific endonuclease BisI (recognition site 5’-G(5mC)↓NGC-3’), but differs in positions of DNA cleavage producing 3’-protruding ends. Due to its ability to cleave only methylated DNA, enzyme BlsI can find an application in gene engineering works as well as in determining the methylation status of eucaryotic DNA.

Read more: Site-specific endonuclease BlsI recognizes DNA sequence 5’-G(5mC)N^GC-3’ and cleaves it producing...

A novel site-specific endonuclease GluI recognizes methylated DNA sequence 5’-G(5mC)^NG(5mC)-3’/ 3’-(5mC)GN^(5mC)G

This email address is being protected from spambots. You need JavaScript enabled to view it. , E.V. Chmuzh, Yu.E. Tomilova, T.N. Nayakshina, D.A. Gonchar, V.S. Dedkov, S.Kh. Degtyarev

Translated from "Ovchinnikov bulletin of biotechnology and physical and chemical biology" V.3, No 2, pp 13-17, 2007

 

A novel site-specific endonuclease GluI from the bacterial strain GL24 has been isolated and characterized. The enzyme recognizes methylated DNA sequence 5’-G(5mC)^NG(5mC)-3’/3’-(5mC)GN^(5mC)G-5’ and cleaves it as it is shown by arrow. Due to its ability to cleave only modified DNA GluI may be useful for genetic engineering experiments as well as for determination of DNA methylation status in eucaryotes.

Read more: A novel site-specific endonuclease GluI recognizes methylated DNA sequence 5’-G(5mC)^NG(5mC)-3’/...

New Eight Bases Cutter AbsI from Arthrobacter species Recognizes Palindromic DNA Sequence 5’-CC^TCGAGG-3’

This email address is being protected from spambots. You need JavaScript enabled to view it. , Yu.G. Kashirina, Yu.E. Tomilova, D.A. Gonchar, V.S. Dedkov, N.A. Mikhnenkova, S.Kh. Degtryarev

Translated from "Ovchinnikov bulletin of biotechnology and physical and chemical biology" V.2, No 2, pp 29-34, 2006

 

Bacterial strain Arthrobacter species7M06 producing site-specific endonuclease AbsI has been discovered. AbsI recognizes palindromic octanucleotide DNA sequence 5’-CC↑TCGAGG-3’ and hydrolyzes it after second cytosine, producing 5’- sticky ends, which are compatible with sticky ends after DNA cleavage by restriction endonucleases XhoI (5’-C↑TCGAG-3’), PspXI (5’-VC↑TCGAGB-3’) and SalI (5’-G↑TCGAC-3’). Among all known rare-cutting site-specific endonucleases AbsI is the only enzyme which has no recognition sequences in standard substrates Lambda and T7 DNAs and Adenovirus type 2 DNA

Read more: New Eight Bases Cutter AbsI from Arthrobacter species Recognizes Palindromic DNA Sequence...

A novel restriction endonuclease GlaI recognizes methylated sequence 5’-G(5mC)^GC-3’

This email address is being protected from spambots. You need JavaScript enabled to view it. , Tatyana N. Najakshina, Murat A. Abdurashitov, Julia E. Tomilova, Nina V. Mezentzeva, Vladimir S. Dedkov, Natalya A. Mikhnenkova, Danila A. Gonchar, Sergei Kh. Degtyarev

Translated from Biotechnologia (russ.). 2006. N 4. P. 31-35

 

A novel restriction endonuclease GlaI from the bacterial strain GL29 has been isolated and characterized. The enzyme recognizes methylated DNA sequence 5’-G(5mC)↓GC-3’ and cleaves it as indicated by arrow. Due to its ability to cleave only modified DNA GlaI may find a practical application in genetic engineering experiments as well as in determination of DNA methylation status

Read more: A novel restriction endonuclease GlaI recognizes methylated sequence 5’-G(5mC)^GC-3’