SibEnzyme REBase

restriction enzymes database

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Gla I

Available supply options

Enzyme name

Gla I

Prototype

GlaI

SKU

SE-E493

Turbo version

Not available

High-concentration version

Not available

Restriction type

Type IIMi, Type IIPi

Recognition site

5'… R(5mC)^▼GY …3'

3'… YG_▲(5mC)R …5'

Source

An E.coli strain that carries the cloned Gla I gene from Glacial ice bacterium Gl29

Optimal buffer

SE-buffer Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.)

Optimal temperature

30 °C

Inactivation temperature

65 °C

Buffer activity

B	G	O	W	Y	ROSE
75	75	25	25	100	100

Unit definition

One unit is defined as the amount of enzyme required to hydrolyse completely a unique 5`-G(5mC)G(5mC)-3`/3`-(5mC)G(5mC)G-5` site in 1 μg of pHspAI2 plasmid DNA, which is linearized with GsaI, in 1 hour at 30°C in a total reaction volume of 50 μl. As a result of this site hydrolysis two main DNA fragments (3,419 and 699 bp) are produced (see lanes 3-5 in the figure). GlaI digestion of recognition sequences with three and two 5-methylcytosines results in additional bands appearance (lane 6 in the figure). GlaI activity assay on DNA pHspAI2/GsaI Lanes: 2 Control pHspAI2/GsaI DNA 3 - 0.5 μl GlaI (diluted 1/100), 4 - 1 μl GlaI (diluted 1/100), 5 - 2 μl GlaI (diluted 1/100), 6 - 1 μl of undiluted GlaI, 1 and 7- 1 Kb SE DNA Ladder.Products were separated in 1% agarose gel in TAE Buffer. In the presence of 20% DMSO, the enzyme's activity significantly increases without losing specificity

Assayed on

DNA pHspAI2/GsaI is a linearized plasmid pHspAI2, which carries a gene of DNA-methyltransferase M.HspAI (recognition sequence 5`-GCGC-3`) and includes a unique GlaI recognition site 5`-G(5mC)G(5mC)-3`/3`-(5mC)G(5mC)G-5` [2].

Storage conditions

10 mM Tris-HCl (pH 7.6); 250 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 0,1% Triton X-100, 0.05 mg/ml BSA, 50% glycerol; Store at -20°C.

Ligation

Nonspecific hydrolysis

No detectable degradation of 1μg of Lambda DNA was observed after incubation with 100 units of enzyme for 16 hours at 30°C in a total reaction volume of 50 μl.

Methylation sensitivity

The enzyme cleaves only C5-methylated DNA and does not cut unmodified DNAand DNA with N4-methylcytosines [1].

Reagents supplied with enzyme

10 X SE-buffer Y, pHspAI2/GsaI DNA

Notes

References

M.A. Abdurashitov, A.G. Akishev, S.Kh. Degtyarev Determination of the Methylation Status of RCGY Sites in the Human Genome: A Comparison of NGS Results from GlaI Hydrolysates and Data from GlaI-PCR Analysis // Epigenetic DNA Diagnostics, Vol. 2025(1), DOI: 10.26213/3034-4298.2025.6.1.001 (2025). (In Russian) 2025
External publication L. Zhen, X. Tang, Z. Xu, Y. Huang, X. Qian, H. Lin, C. Li, R. Cui, H. Fang, H. Yang, J. Qiu, Z. Fang, X. Peng, Y. Jin, J. Nie, S. Guo, Y. Wang, M. Zhong, H. Gu, H. Xu Early Diagnosis of Colorectal Cancer Based on Bisulfite-free Site-specific Methylation Identification PCR Strategy: High-Sensitivity, Accuracy, and Primary Medical Accessibility. // Adv. Sci. 2024, 2401137. 2024
External publication Qiaomin Wu, Yang Yu, Mengqi Chen, Jinyan Long, Xiaolan Yang A label-free fluorescence sensing strategy based on GlaI-assisted EXPAR for rapid and accurate quantification of human methyltranferase activity // Talanta, 269, 125456 (2024) 2024
External publication Xu, G., Yang, H., Qiu, J. et al. Sequence terminus dependent PCR for site-specific mutation and modification detection. // Nature Communications 14, 1169 (2023) 2023
A.G. Akishev, N.A. Netesova, M.A. Abdurashitov, S.Kh. Degtyarev Determination of G/C polymorphism at position chr20:37352001 in human blood DNA samples using GlaI- and Bst2UI-PCR analysis, Genetics. 2023. Vol. 59, No. 11. pp. 1303–1312, DOI: 10.31857/S0016675823110012 (2023). (In Russian) 2023
External publication Zhou S, Sun H, Huo D, Wang X, Qi N, Peng L, Yang M, Lu P, Hou C. A novel methyl-dependent DNA endonuclease GlaI coupling with double cascaded strand displacement amplification and CRISPR/Cas12a for ultra-sensitive detection of DNA methylation. // Analytica Chimica Acta, Volume 1212, 15 June 2023 2022
External publication Dong N, Wang W, Qin Y, Wang Y, Shan H. Sensitive lateral flow assay for bisulfite-free DNA methylation detection based on the restriction endonuclease GlaI and rolling circle amplification // Analytica Chimica Acta, Volume 1227, 22 September 2023 2022
A.G. Akishev, N.A. Netesova, M.A. Abdurashitov, S.Kh. Degtyarev Detection of the 5mC/T polymorphism at position Chr1:245618129 in human blood DNA samples using GlaI- and FatI-PCR analysis // Epigenetic DNA Diagnostics, DOI: 10.26213/SE.2019.69.42836 (2021). (In Russian) 2021
A.G. Akishev, N.A. Netesova, M.A. Abdurashitov, I.V. Vikhlyanov, M.K. Nikitin, A.B. Karpov, S.Kh. Degtyarev Assessment of the Risk of Breast and Stomach Cancer by Determining the 5mC/T Polymorphism Variants in the KIF26B Gene Intron at Position chr1:245618129 // Epigenetic DNA Diagnostics, DOI: 10.26213/SE.2019.78.40307 (2021). (In Russian) 2021
A.G. Akishev, N.A. Netesova, M.A. Abdurashitov, S.Kh. Degtyarev Detection of the 5mC/T polymorphism in the AluSx repeat (CHR16: 75033884) in human blood DNA preparations using GlaI- and FatI-PCR analysis // Epigenetic DNA Diagnostics, DOI: 10.26213/SE.2021.11.76.001 (2021). (In Russian) 2021
A.G. Akishev, N.A. Netesova, M.A. Abdurashitov, S.Kh. Degtyarev Investigation of RCGY Site Methylation in the Regulatory Region of the FAM19A4 Gene in Human Cell Line DNA Preparations Using GLAD-PCR Analysis // Epigenetic DNA Diagnostics, DOI: 10.26213/SE.2021.97.62.001 (2021). (In Russian) 2021
B.S. Malyshev, N.A. Netesova, N.A. Smetannikova, M.A. Abdurashitov, A.G. Akishev, E.V. Dubinin, A.Z. Azanov, I.V. Vikhlyanov, M.K. Nikitin, A.B. Karpov, S.Kh. Degtyarev GlaD-PCR Analysis of DNA Methylation Sites in Regulatory Regions of Tumor Suppressor Genes in Gastric Cancer // Acta Naturae, 2020, Vol. 12, No. 3(46): 124–133 (2020). (In Russian) 2020
External publication Petrova, D. V., Naumenko, M. B., Khantakova, D. V., Grin, I. R. & Zharkov, D. O. Relative Efficiency of Recognition of 5-Methylcytosine and 5-Hydroxymethylcytosine by Methyl-Dependent DNA Endonuclease GlaI // Russian Journal of Bioorganic Chemistry T. 45, № 6, стр. 625-629. (2019) 2019
V.A. Chernukhin, V.S. Dedkov, D.A. Gonchar, M.A. Abdurashitov, A.G. Akishev, T.N. Nayakshina, E.N. Lomakovskaya, S.Kh. Degtyarev Comparison of the substrate specificity of GlaI recombinant site-specific methyl-directed DNA endonuclease and the native enzyme isolated from Glacial ice bacterium strain // DNA processing enzymes , DOI: 10.26213/SE.2019.81.28075, (2019). (in Russian) 2019
A.G. Akishev, N.A. Netesova, M.A. Abdurashitov, I.V. Vikhlyanov, M.K. Nikitin, A.B. Karpov, S.Kh. Degtyarev GlaI-PCR analysis of methylation of ACGC site in Chr11: 65647266 in DNA samples from the blood cells of healthy donors and early stage breast cancers // Epigenetic DNA diagnostics, DOI: 10.26213/SE.2019.66.12.001, (2019). 2019
N.A Smetannikova., A.A. Evdokimov, N.A. Netesova, M.A. Abdurashitov, A.G. Akishev, E.V. Dubinin, P.I. Pozdnyakov, I.V. Vihlyanov, M.K. Nikitin, E.B. Topolnitsky, A.B. Karpov, S.A. Kolomiets, S.K. Degtyarev Application of GLAD-PCR Assay for Study on DNA Methylation in Regulatory Regions of Some Tumor-Suppressor Genes in Lung Cancer // Zhongguo Fei Ai Za Zhi. 2019 Sep 20;22(9):551-561. 2019
M.A. Abdurashitov, V.N. Tomilov, D.A. Gonchar, A.V. Snezhkina, G.S. Krasnov, A.V. Kudryavtseva, S.Kh. Degtyarev Comparative analysis of RCGY sites methylation in three human cell lines. // Epigenetic DNA diagnoctics, vol.2019(1), DOI: 10.26213/SE.2019. 76.40116 2019
External publication Susan M Mitchell, Keith N Rand, Zheng-Zhou Xu, Thu Ho, Glenn S Brown, Jason P Ross, Peter L Molloy Helper-Dependent Chain Reaction (HDCR) for Selective Amplification of Methylated DNA Sequences // Methods Mol Biol (2018) 2018
External publication Yueying Sun, Yuanyuan Sun, Weimin Tian, Chenghui Liu, * Kejian Gao and Zhengping L A novel restriction endonuclease GlaI for rapid and highly sensitive detection of DNA methylation coupled with isothermal exponential amplification reaction // Chemical Science, 9, pp.1344-1351 (2018) 2018
Tomilov V.N., Abdurashitov M.A., Gonchar D.A., Snezhkina A.V., Krasnov G.S., Kudryavtseva A.V., Degtyarev S.Kh. Comparative analysis of RCGY sites methylation in the genomes of Raji and U-937 malignant and normal lung fibroblast cell lines // Conference papers: Cancer Epigenetics and Biomarkers, Osaka, Japan, October 2017. 2017
A.G. Akishev et al. GLAD-PCR assay of selected R(5mC)GY sites in URB1 and CEPBD genes in human genome // Res J Pharm Biol Chem Sci, 8(1): pp.465-475, 2017. 2017
M.A. Abdurashitov , V.A. Chernukhin , D.A. Gonchar , V.S. Dedkov, N.A. Mikhnenkova, S.Kh. Degtyarev Dimethyl Sulfoxide Changes the Recognition Site Preference of Methyl- directed Site-specific DNA Endonuclease GlaI. // Research Journal of Pharmaceutical, Biological and Chemical Sciences, vol. 7(1),p. 1733-1739 (2016). 2016
E.V. Dubinin, A.G. Akishev, M.A. Abdurashitov, S.B. Oleynikova, V.L. Sitko, S.Kh. Degtyarev Real time GlaI-PCR assay of regulation regions of human genes HDAC4, RARB and URB1 // // Research Journal of Pharmaceutical, Biological and Chemical Sciences, vol 7(2), p.p. 667-676 (2016). 2016
A.A. Evdokimov, N.A. Netesova, N.A. Smetannikova, M.A. Abdurashitov, A.G. Akishev, E.S. Davidovich, V.V. Kuznetsov, Yu.D. Ermolaev, A.B. Karpov, A.E. Sazonov, R.M. Takhaouov, S.Kh. Degtyarev Application of GLAD-PCR assay for determination of the methylation sites in the regulatory regions of tumor-supressors gene ELMO1 and ESR1 in colorectal cancer // Translated from Problems in oncology, #1, p.116-120 (2016). (In Russian) 2016
A.A. Evdokimov et al. GLAD-PCR Assay of DNA Methylation Markers Associated with Colorectal Cancer // Biol Med (Aligarh) , 8:7(2016). 2016
M.A. Abdurashitov, V.N. Tomilov, D.A. Gonchar, V.V. Kuznetsov, S.Kh. Degtyarev Mapping of R(5mC)GY Sites in the Genome of Human Malignant Cell Line Raji // Biol Med (Aligarh) Volume 7, Issue 4, BM-135-15. (2015). 2015
V.V. Kuznetsov, M.A. Abdurashitov, A.G. Akishev, S.Kh. Degtyarev Method of determining nucleotide sequence Pu(5mC)GPy at predetermined position of long-distance DNA. // Russian Federation patent RU 2525710 C1 (2014). (In Russian) 2014
External publication Fagan RL, Wu M, Chedin F, Brenner C An Ultrasensitive High Throughput Screen for DNA Methyltransferase 1-Targeted Molecular Probes // PLoS ONE 8(11): e78752. doi:10.1371/journal.pone.0078752 (2013) 2013
E.V. Zemlyanskaya, S.Kh. Degtyarev Substrate specificity and properties of methyl-directed site-specific DNA endonucleases // Molecular Biology, v.47, No.6, p.900-913 (2013). (In Russian) 2013
M.A. Abdurashitov, A.N. Kuksova, A.G. Akishev, E.V. Zemlyanskaya, S.Kh. Degtyarev MteI-PCR assay - a new method of DNA methylation study in GC rich regulation regions of human tumor supressor genes // Bulletin of biotechnology and physico-chemical biology named by Yu.A.Ovchinnikov (Moscow), V.9, No.3, p.15-23 (2013). (In Russian) 2013
External publication Keith N. Rand, Graeme P. Young, Thu Ho and Peter L. Molloy Sensitive and selective amplification of methylated DNA sequences using helper-dependent chain reaction in combination with a methylationdependent restriction enzymes. // Nucleic Acids Research, pp. 1-10 (2012). 2012
External publication F. Syeda, R.L. Fagan, M. Wean, G.V. Awakumov, J.R. Walker, S. Xue, S. Dhe-Paganon, & C. Brenner The RFTS Domain is a DNA-competitive Inhibitor of Dnmt1 // , JBC, v. 286, pp. 15344-15351 (2011). 2011
A.G. Akishev, D.A. Gonchar, M.A. Abdurashitov, S.Kh. Degtyarev Epigenetic typing of human cancer cell lines by BlsI- and GlaI-PCR assays. // Ovchinnikov bulletin of biotechnology and physical and chemical biology V.7, No 3, pp 5-16, 2011. 2011
External publication Kravets AP, Mousseau TA, Litvinchuk AV, Ostermiller Sh, Vengen GS, Grodzinski DM Changes in wheat DNA methylation pattern after chronic seed gamma-irradiation. // Tsitol Genet. 2010 Sep-Oct;44(5):18-22. Russian. 2010
D.A. Gonchar, A.G. Akishev, S.Kh. Degtyarev BlsI- and GlaI-PCR assays – a new method of DNA methylation study // Ovchinnikov bulletin of biotechnology and physical and chemical biology , V.6, No 1, p. 5-12 (2010). (in Russian) 2010
M.A. Abdurashitov, V.A. Chernukhin, D.A. Gonchar, S.Kh. Degtyarev GlaI digestion of mouse γ-satellite DNA: study of primary structure and ACGT sites methylation // BMC Genomics 2009, 10:322. 2009
D.A. Gonchar, A.G. Akishev, S.Kh. Degtyarev A method for identifying hypermethylated CpG islands in the regions of tumour-suppressor genes in human DNA. // Patent RU 2413773 C1 (2009). (In Russian) 2009
G.V. Tarasova, T.N. Nayakshina, S.Kh. Degtyarev Substrate specificity of new methyl-directed DNA endonuclease GlaI . // BMC Molecular Biology 2008, 9:7. 2008
V.A. Chernukhin, M.A. Abdurashitov, V.N. Tomilov, D.A. Gonchar, S.Kh. Degtyarev Comparative analysis of mouse chromosomal DNA digestion with restriction endonucleases in vitro and in silico // Ovchinnikov bulletin of biotechnology and physical and chemical biology V.3, No 4, p. 19-27.(2007) (In Russian) 2007
V.A. Chernukhin, T.N. Nayakshina, M.A. Abdurashitov, Y.E. Tomilova, N.V. Mezentseva, V.S. Dedkov, N.A. Mikhnenkova, D.A. Gonchar, S.Kh. Degtyarev A novel restriction endonuclease GlaI recognizes methylated sequence 5′-G(5mC)^GC-3′. // Biotechnologia V 4. P. 31-35(2006). (In Russian) 2006
Y.E. Tomilova, V.A. Chernukhin, S.Kh. Degtyarev Dependence of site-specific endonuclease GlaI activity on quantity and location of methylcytosines in the recognition sequence 5`-GCGC-3`. // Bulletin of biotechnology and physico-chemical biology V.2, No 1, p. 30-39 (2006). (In Russian) 2006
V.A. Chernukhin, T.N. Nayakshina, N.V. Mezentseva, Y.E. Tomilova, S.Kh. Degtyarev, V.S. Dedkov Bacterial strain Glacial ice bacterium I - producer of GlaI restriction endonuclease. // Russian Federation patent RU 2287012 C1 (2005). (In Russian) 2005
V.E. Repin, E.A. Prikhodko, S.Kh. Degtyarev Search of strains producers of restriction endonucleases among soil microorganisms. // Izvestia SB SA USSR, Biological sciences series, No.2, 109-112 (1988). (In Russian) 1988

Sequence search

Enzyme list

← Back to list

Gla I

Available supply options

Enzyme name

Gla I

Prototype

GlaI

SKU

SE-E493

Turbo version

Not available

High-concentration version

Not available

Restriction type

Type IIMi, Type IIPi

Recognition site

5'… R(5mC)^▼GY …3'

3'… YG_▲(5mC)R …5'

Source

An E.coli strain that carries the cloned Gla I gene from Glacial ice bacterium Gl29

Optimal buffer

SE-buffer Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.)

Optimal temperature

30 °C

Inactivation temperature

65 °C

Buffer activity

B	G	O	W	Y	ROSE
75	75	25	25	100	100

Unit definition

Assayed on

Storage conditions

10 mM Tris-HCl (pH 7.6); 250 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 0,1% Triton X-100, 0.05 mg/ml BSA, 50% glycerol; Store at -20°C.

Ligation

Nonspecific hydrolysis

No detectable degradation of 1μg of Lambda DNA was observed after incubation with 100 units of enzyme for 16 hours at 30°C in a total reaction volume of 50 μl.

Methylation sensitivity

The enzyme cleaves only C5-methylated DNA and does not cut unmodified DNAand DNA with N4-methylcytosines [1].

Reagents supplied with enzyme

10 X SE-buffer Y, pHspAI2/GsaI DNA

Notes

References

M.A. Abdurashitov, A.G. Akishev, S.Kh. Degtyarev Determination of the Methylation Status of RCGY Sites in the Human Genome: A Comparison of NGS Results from GlaI Hydrolysates and Data from GlaI-PCR Analysis // Epigenetic DNA Diagnostics, Vol. 2025(1), DOI: 10.26213/3034-4298.2025.6.1.001 (2025). (In Russian) 2025
External publication L. Zhen, X. Tang, Z. Xu, Y. Huang, X. Qian, H. Lin, C. Li, R. Cui, H. Fang, H. Yang, J. Qiu, Z. Fang, X. Peng, Y. Jin, J. Nie, S. Guo, Y. Wang, M. Zhong, H. Gu, H. Xu Early Diagnosis of Colorectal Cancer Based on Bisulfite-free Site-specific Methylation Identification PCR Strategy: High-Sensitivity, Accuracy, and Primary Medical Accessibility. // Adv. Sci. 2024, 2401137. 2024
External publication Qiaomin Wu, Yang Yu, Mengqi Chen, Jinyan Long, Xiaolan Yang A label-free fluorescence sensing strategy based on GlaI-assisted EXPAR for rapid and accurate quantification of human methyltranferase activity // Talanta, 269, 125456 (2024) 2024
External publication Xu, G., Yang, H., Qiu, J. et al. Sequence terminus dependent PCR for site-specific mutation and modification detection. // Nature Communications 14, 1169 (2023) 2023
A.G. Akishev, N.A. Netesova, M.A. Abdurashitov, S.Kh. Degtyarev Determination of G/C polymorphism at position chr20:37352001 in human blood DNA samples using GlaI- and Bst2UI-PCR analysis, Genetics. 2023. Vol. 59, No. 11. pp. 1303–1312, DOI: 10.31857/S0016675823110012 (2023). (In Russian) 2023
External publication Zhou S, Sun H, Huo D, Wang X, Qi N, Peng L, Yang M, Lu P, Hou C. A novel methyl-dependent DNA endonuclease GlaI coupling with double cascaded strand displacement amplification and CRISPR/Cas12a for ultra-sensitive detection of DNA methylation. // Analytica Chimica Acta, Volume 1212, 15 June 2023 2022
External publication Dong N, Wang W, Qin Y, Wang Y, Shan H. Sensitive lateral flow assay for bisulfite-free DNA methylation detection based on the restriction endonuclease GlaI and rolling circle amplification // Analytica Chimica Acta, Volume 1227, 22 September 2023 2022
A.G. Akishev, N.A. Netesova, M.A. Abdurashitov, S.Kh. Degtyarev Detection of the 5mC/T polymorphism at position Chr1:245618129 in human blood DNA samples using GlaI- and FatI-PCR analysis // Epigenetic DNA Diagnostics, DOI: 10.26213/SE.2019.69.42836 (2021). (In Russian) 2021
A.G. Akishev, N.A. Netesova, M.A. Abdurashitov, I.V. Vikhlyanov, M.K. Nikitin, A.B. Karpov, S.Kh. Degtyarev Assessment of the Risk of Breast and Stomach Cancer by Determining the 5mC/T Polymorphism Variants in the KIF26B Gene Intron at Position chr1:245618129 // Epigenetic DNA Diagnostics, DOI: 10.26213/SE.2019.78.40307 (2021). (In Russian) 2021
A.G. Akishev, N.A. Netesova, M.A. Abdurashitov, S.Kh. Degtyarev Detection of the 5mC/T polymorphism in the AluSx repeat (CHR16: 75033884) in human blood DNA preparations using GlaI- and FatI-PCR analysis // Epigenetic DNA Diagnostics, DOI: 10.26213/SE.2021.11.76.001 (2021). (In Russian) 2021
A.G. Akishev, N.A. Netesova, M.A. Abdurashitov, S.Kh. Degtyarev Investigation of RCGY Site Methylation in the Regulatory Region of the FAM19A4 Gene in Human Cell Line DNA Preparations Using GLAD-PCR Analysis // Epigenetic DNA Diagnostics, DOI: 10.26213/SE.2021.97.62.001 (2021). (In Russian) 2021
B.S. Malyshev, N.A. Netesova, N.A. Smetannikova, M.A. Abdurashitov, A.G. Akishev, E.V. Dubinin, A.Z. Azanov, I.V. Vikhlyanov, M.K. Nikitin, A.B. Karpov, S.Kh. Degtyarev GlaD-PCR Analysis of DNA Methylation Sites in Regulatory Regions of Tumor Suppressor Genes in Gastric Cancer // Acta Naturae, 2020, Vol. 12, No. 3(46): 124–133 (2020). (In Russian) 2020
External publication Petrova, D. V., Naumenko, M. B., Khantakova, D. V., Grin, I. R. & Zharkov, D. O. Relative Efficiency of Recognition of 5-Methylcytosine and 5-Hydroxymethylcytosine by Methyl-Dependent DNA Endonuclease GlaI // Russian Journal of Bioorganic Chemistry T. 45, № 6, стр. 625-629. (2019) 2019
V.A. Chernukhin, V.S. Dedkov, D.A. Gonchar, M.A. Abdurashitov, A.G. Akishev, T.N. Nayakshina, E.N. Lomakovskaya, S.Kh. Degtyarev Comparison of the substrate specificity of GlaI recombinant site-specific methyl-directed DNA endonuclease and the native enzyme isolated from Glacial ice bacterium strain // DNA processing enzymes , DOI: 10.26213/SE.2019.81.28075, (2019). (in Russian) 2019
A.G. Akishev, N.A. Netesova, M.A. Abdurashitov, I.V. Vikhlyanov, M.K. Nikitin, A.B. Karpov, S.Kh. Degtyarev GlaI-PCR analysis of methylation of ACGC site in Chr11: 65647266 in DNA samples from the blood cells of healthy donors and early stage breast cancers // Epigenetic DNA diagnostics, DOI: 10.26213/SE.2019.66.12.001, (2019). 2019
N.A Smetannikova., A.A. Evdokimov, N.A. Netesova, M.A. Abdurashitov, A.G. Akishev, E.V. Dubinin, P.I. Pozdnyakov, I.V. Vihlyanov, M.K. Nikitin, E.B. Topolnitsky, A.B. Karpov, S.A. Kolomiets, S.K. Degtyarev Application of GLAD-PCR Assay for Study on DNA Methylation in Regulatory Regions of Some Tumor-Suppressor Genes in Lung Cancer // Zhongguo Fei Ai Za Zhi. 2019 Sep 20;22(9):551-561. 2019
M.A. Abdurashitov, V.N. Tomilov, D.A. Gonchar, A.V. Snezhkina, G.S. Krasnov, A.V. Kudryavtseva, S.Kh. Degtyarev Comparative analysis of RCGY sites methylation in three human cell lines. // Epigenetic DNA diagnoctics, vol.2019(1), DOI: 10.26213/SE.2019. 76.40116 2019
External publication Susan M Mitchell, Keith N Rand, Zheng-Zhou Xu, Thu Ho, Glenn S Brown, Jason P Ross, Peter L Molloy Helper-Dependent Chain Reaction (HDCR) for Selective Amplification of Methylated DNA Sequences // Methods Mol Biol (2018) 2018
External publication Yueying Sun, Yuanyuan Sun, Weimin Tian, Chenghui Liu, * Kejian Gao and Zhengping L A novel restriction endonuclease GlaI for rapid and highly sensitive detection of DNA methylation coupled with isothermal exponential amplification reaction // Chemical Science, 9, pp.1344-1351 (2018) 2018
Tomilov V.N., Abdurashitov M.A., Gonchar D.A., Snezhkina A.V., Krasnov G.S., Kudryavtseva A.V., Degtyarev S.Kh. Comparative analysis of RCGY sites methylation in the genomes of Raji and U-937 malignant and normal lung fibroblast cell lines // Conference papers: Cancer Epigenetics and Biomarkers, Osaka, Japan, October 2017. 2017
A.G. Akishev et al. GLAD-PCR assay of selected R(5mC)GY sites in URB1 and CEPBD genes in human genome // Res J Pharm Biol Chem Sci, 8(1): pp.465-475, 2017. 2017
M.A. Abdurashitov , V.A. Chernukhin , D.A. Gonchar , V.S. Dedkov, N.A. Mikhnenkova, S.Kh. Degtyarev Dimethyl Sulfoxide Changes the Recognition Site Preference of Methyl- directed Site-specific DNA Endonuclease GlaI. // Research Journal of Pharmaceutical, Biological and Chemical Sciences, vol. 7(1),p. 1733-1739 (2016). 2016
E.V. Dubinin, A.G. Akishev, M.A. Abdurashitov, S.B. Oleynikova, V.L. Sitko, S.Kh. Degtyarev Real time GlaI-PCR assay of regulation regions of human genes HDAC4, RARB and URB1 // // Research Journal of Pharmaceutical, Biological and Chemical Sciences, vol 7(2), p.p. 667-676 (2016). 2016
A.A. Evdokimov, N.A. Netesova, N.A. Smetannikova, M.A. Abdurashitov, A.G. Akishev, E.S. Davidovich, V.V. Kuznetsov, Yu.D. Ermolaev, A.B. Karpov, A.E. Sazonov, R.M. Takhaouov, S.Kh. Degtyarev Application of GLAD-PCR assay for determination of the methylation sites in the regulatory regions of tumor-supressors gene ELMO1 and ESR1 in colorectal cancer // Translated from Problems in oncology, #1, p.116-120 (2016). (In Russian) 2016
A.A. Evdokimov et al. GLAD-PCR Assay of DNA Methylation Markers Associated with Colorectal Cancer // Biol Med (Aligarh) , 8:7(2016). 2016
M.A. Abdurashitov, V.N. Tomilov, D.A. Gonchar, V.V. Kuznetsov, S.Kh. Degtyarev Mapping of R(5mC)GY Sites in the Genome of Human Malignant Cell Line Raji // Biol Med (Aligarh) Volume 7, Issue 4, BM-135-15. (2015). 2015
V.V. Kuznetsov, M.A. Abdurashitov, A.G. Akishev, S.Kh. Degtyarev Method of determining nucleotide sequence Pu(5mC)GPy at predetermined position of long-distance DNA. // Russian Federation patent RU 2525710 C1 (2014). (In Russian) 2014
External publication Fagan RL, Wu M, Chedin F, Brenner C An Ultrasensitive High Throughput Screen for DNA Methyltransferase 1-Targeted Molecular Probes // PLoS ONE 8(11): e78752. doi:10.1371/journal.pone.0078752 (2013) 2013
E.V. Zemlyanskaya, S.Kh. Degtyarev Substrate specificity and properties of methyl-directed site-specific DNA endonucleases // Molecular Biology, v.47, No.6, p.900-913 (2013). (In Russian) 2013
M.A. Abdurashitov, A.N. Kuksova, A.G. Akishev, E.V. Zemlyanskaya, S.Kh. Degtyarev MteI-PCR assay - a new method of DNA methylation study in GC rich regulation regions of human tumor supressor genes // Bulletin of biotechnology and physico-chemical biology named by Yu.A.Ovchinnikov (Moscow), V.9, No.3, p.15-23 (2013). (In Russian) 2013
External publication Keith N. Rand, Graeme P. Young, Thu Ho and Peter L. Molloy Sensitive and selective amplification of methylated DNA sequences using helper-dependent chain reaction in combination with a methylationdependent restriction enzymes. // Nucleic Acids Research, pp. 1-10 (2012). 2012
External publication F. Syeda, R.L. Fagan, M. Wean, G.V. Awakumov, J.R. Walker, S. Xue, S. Dhe-Paganon, & C. Brenner The RFTS Domain is a DNA-competitive Inhibitor of Dnmt1 // , JBC, v. 286, pp. 15344-15351 (2011). 2011
A.G. Akishev, D.A. Gonchar, M.A. Abdurashitov, S.Kh. Degtyarev Epigenetic typing of human cancer cell lines by BlsI- and GlaI-PCR assays. // Ovchinnikov bulletin of biotechnology and physical and chemical biology V.7, No 3, pp 5-16, 2011. 2011
External publication Kravets AP, Mousseau TA, Litvinchuk AV, Ostermiller Sh, Vengen GS, Grodzinski DM Changes in wheat DNA methylation pattern after chronic seed gamma-irradiation. // Tsitol Genet. 2010 Sep-Oct;44(5):18-22. Russian. 2010
D.A. Gonchar, A.G. Akishev, S.Kh. Degtyarev BlsI- and GlaI-PCR assays – a new method of DNA methylation study // Ovchinnikov bulletin of biotechnology and physical and chemical biology , V.6, No 1, p. 5-12 (2010). (in Russian) 2010
M.A. Abdurashitov, V.A. Chernukhin, D.A. Gonchar, S.Kh. Degtyarev GlaI digestion of mouse γ-satellite DNA: study of primary structure and ACGT sites methylation // BMC Genomics 2009, 10:322. 2009
D.A. Gonchar, A.G. Akishev, S.Kh. Degtyarev A method for identifying hypermethylated CpG islands in the regions of tumour-suppressor genes in human DNA. // Patent RU 2413773 C1 (2009). (In Russian) 2009
G.V. Tarasova, T.N. Nayakshina, S.Kh. Degtyarev Substrate specificity of new methyl-directed DNA endonuclease GlaI . // BMC Molecular Biology 2008, 9:7. 2008
V.A. Chernukhin, M.A. Abdurashitov, V.N. Tomilov, D.A. Gonchar, S.Kh. Degtyarev Comparative analysis of mouse chromosomal DNA digestion with restriction endonucleases in vitro and in silico // Ovchinnikov bulletin of biotechnology and physical and chemical biology V.3, No 4, p. 19-27.(2007) (In Russian) 2007
V.A. Chernukhin, T.N. Nayakshina, M.A. Abdurashitov, Y.E. Tomilova, N.V. Mezentseva, V.S. Dedkov, N.A. Mikhnenkova, D.A. Gonchar, S.Kh. Degtyarev A novel restriction endonuclease GlaI recognizes methylated sequence 5′-G(5mC)^GC-3′. // Biotechnologia V 4. P. 31-35(2006). (In Russian) 2006
Y.E. Tomilova, V.A. Chernukhin, S.Kh. Degtyarev Dependence of site-specific endonuclease GlaI activity on quantity and location of methylcytosines in the recognition sequence 5`-GCGC-3`. // Bulletin of biotechnology and physico-chemical biology V.2, No 1, p. 30-39 (2006). (In Russian) 2006
V.A. Chernukhin, T.N. Nayakshina, N.V. Mezentseva, Y.E. Tomilova, S.Kh. Degtyarev, V.S. Dedkov Bacterial strain Glacial ice bacterium I - producer of GlaI restriction endonuclease. // Russian Federation patent RU 2287012 C1 (2005). (In Russian) 2005
V.E. Repin, E.A. Prikhodko, S.Kh. Degtyarev Search of strains producers of restriction endonucleases among soil microorganisms. // Izvestia SB SA USSR, Biological sciences series, No.2, 109-112 (1988). (In Russian) 1988

Isoshizomers

Isoschizomers and neoschizomers

SE enzyme	Recognition site	Known analogs	Type	Recognition site
Aat II	5'… GACGT^▼C …3' 3'… C_▲TGCAG …5'	Zra I	Neoschizomer	5'… GAC^▼GTC …3' 3'… CTG_▲CAG …5'
Acc16 I	5'… TGC^▼GCA …3' 3'… ACG_▲CGT …5'	AviII FspI MstI NsbI	Isoschizomer	5'… TGC^▼GCA …3' 3'… ACG_▲CGT …5'
Acc36 I	5'… ACCTGC(N)₄^▼ …3' 3'… TGGACG(N)₈_▲ …5'	BfuAI BspMI BveI	Isoschizomer	5'… ACCTGC(N)₄^▼ …3' 3'… TGGACG(N)₈_▲ …5'
Acc65 I	5'… G^▼GTACC …3' 3'… CCATG_▲G …5'	Asp718I	Isoschizomer	5'… G^▼GTACC …3' 3'… CCATG_▲G …5'
Acc65 I	5'… G^▼GTACC …3' 3'… CCATG_▲G …5'	Kpn I	Neoschizomer	5'… GGTAC^▼C …3' 3'… C_▲CATGG …5'
AccB1 I	5'… G^▼GYRCC …3' 3'… CCRYG_▲G …5'	BanI BshNI BspT107I HgiCI	Isoschizomer	5'… G^▼GYRCC …3' 3'… CCRYG_▲G …5'
AccB7 I	5'… CCANNNN^▼NTGG …3' 3'… GGTN_▲NNNNACC …5'	PflBI PflMI Van91I	Isoschizomer	5'… CCANNNN^▼NTGG …3' 3'… GGTN_▲NNNNACC …5'
AccBS I	5'… GAG^▼CGG …3' 3'… CTC_▲GCC …5'	BsrBI MbiI	Isoschizomer	5'… GAG^▼CGG …3' 3'… CTC_▲GCC …5'
Acl I	5'… AA^▼CGTT …3' 3'… TTGC_▲AA …5'	Psp1406I	Isoschizomer	5'… AA^▼CGTT …3' 3'… TTGC_▲AA …5'
AclW I	5'… GGATC(N)₄^▼ …3' 3'… CCTAG(N)₅_▲ …5'	AlwI BinI BspPI	Isoschizomer	5'… GGATC(N)₄^▼ …3' 3'… CCTAG(N)₅_▲ …5'
Aco I	5'… Y^▼GGCCR …3' 3'… RCCGG_▲Y …5'	CfrI EaeI	Isoschizomer	5'… Y^▼GGCCR …3' 3'… RCCGG_▲Y …5'
Acs I	5'… R^▼AATTY …3' 3'… YTTAA_▲R …5'	ApoI XapI	Isoschizomer	5'… R^▼AATTY …3' 3'… YTTAA_▲R …5'
Acu I	5'… CTGAAG(N)₁₆^▼ …3' 3'… GACTTC(N)₁₄_▲ …5'	Eco57I	Isoschizomer	5'… CTGAAG(N)₁₆^▼ …3' 3'… GACTTC(N)₁₄_▲ …5'
Afe I	5'… AGC^▼GCT …3' 3'… TCG_▲CGA …5'	Aor51HI Eco47III FunI	Isoschizomer	5'… AGC^▼GCT …3' 3'… TCG_▲CGA …5'
Ahl I	5'… A^▼CTAGT …3' 3'… TGATC_▲A …5'	BcuI SpeI	Isoschizomer	5'… A^▼CTAGT …3' 3'… TGATC_▲A …5'
Ajn I Psp6 I	5'… ^▼CCWGG …3' 3'… GGWCC_▲ …5'	EcoRII PspGI	Isoschizomer	5'… ^▼CCWGG …3' 3'… GGWCC_▲ …5'
Ajn I Psp6 I	5'… ^▼CCWGG …3' 3'… GGWCC_▲ …5'	BptI BseBI Bst2U I BstNI BstOI MvaI	Neoschizomer	5'… CC^▼WGG …3' 3'… GGW_▲CC …5'
Ama87 I	5'… C^▼YCGRG …3' 3'… GRGCY_▲C …5'	AvaI BsiHKCI BsoBI Eco88I NspIII	Isoschizomer	5'… C^▼YCGRG …3' 3'… GRGCY_▲C …5'
Apa I	5'… GGGCC^▼C …3' 3'… C_▲CCGGG …5'	Bsp120I PspOM I	Neoschizomer	5'… G^▼GGCCC …3' 3'… CCCGG_▲G …5'
AsiG I	5'… A^▼CCGGT …3' 3'… TGGCC_▲A …5'	AgeI BshTI CspAI PinAI	Isoschizomer	5'… A^▼CCGGT …3' 3'… TGGCC_▲A …5'
AsiS I Rga I	5'… GCGAT^▼CGC …3' 3'… CGC_▲TAGCG …5'	SgfI	Isoschizomer	5'… GCGAT^▼CGC …3' 3'… CGC_▲TAGCG …5'
AspA2 I	5'… C^▼CTAGG …3' 3'… GGATC_▲C …5'	AvrII BlnI XmaJI	Isoschizomer	5'… C^▼CTAGG …3' 3'… GGATC_▲C …5'
AspLE I BstHH I	5'… GCG^▼C …3' 3'… C_▲GCG …5'	CfoI HhaI	Isoschizomer	5'… GCG^▼C …3' 3'… C_▲GCG …5'
AspLE I BstHH I	5'… GCG^▼C …3' 3'… C_▲GCG …5'	Hin6I HinP1I HspA I	Neoschizomer	5'… G^▼CGC …3' 3'… CGC_▲G …5'
AspS9 I	5'… G^▼GNCC …3' 3'… CCNG_▲G …5'	AsuI Cfr13I PspPI Sau96I	Isoschizomer	5'… G^▼GNCC …3' 3'… CCNG_▲G …5'
AsuC2 I	5'… CC^▼SGG …3' 3'… GGS_▲CC …5'	BcnI CauII NciI	Isoschizomer	5'… CC^▼SGG …3' 3'… GGS_▲CC …5'
AsuHP I	5'… GGTGA(N)₈^▼ …3' 3'… CCACT(N)₇_▲ …5'	HphI	Isoschizomer	5'… GGTGA(N)₈^▼ …3' 3'… CCACT(N)₇_▲ …5'
AsuNH I	5'… G^▼CTAGC …3' 3'… CGATC_▲G …5'	NheI	Isoschizomer	5'… G^▼CTAGC …3' 3'… CGATC_▲G …5'
AsuNH I	5'… G^▼CTAGC …3' 3'… CGATC_▲G …5'	Bmt I	Neoschizomer	5'… GCTAG^▼C …3' 3'… C_▲GATCG …5'
Bbv12 I	5'… GWGCW^▼C …3' 3'… C_▲WCGWG …5'	Alw21I BsiHKAI HgiAI	Isoschizomer	5'… GWGCW^▼C …3' 3'… C_▲WCGWG …5'
Bme18 I	5'… G^▼GWCC …3' 3'… CCWG_▲G …5'	AvaII Eco47I SinI VpaK11BI	Isoschizomer	5'… G^▼GWCC …3' 3'… CCWG_▲G …5'
Bmu I	5'… ACTGGG(N)₅^▼ …3' 3'… TGACCC(N)₄_▲ …5'	BfiI BmrI	Isoschizomer	5'… ACTGGG(N)₅^▼ …3' 3'… TGACCC(N)₄_▲ …5'
Bpm I	5'… CTGGAG(N)₁₆^▼ …3' 3'… GACCTC(N)₁₄_▲ …5'	GsuI	Isoschizomer	5'… CTGGAG(N)₁₆^▼ …3' 3'… GACCTC(N)₁₄_▲ …5'
Bpu14 I	5'… TT^▼CGAA …3' 3'… AAGC_▲TT …5'	AsuII Bsp119I BspT104I BstBI Csp45I NspV SfuI	Isoschizomer	5'… TT^▼CGAA …3' 3'… AAGC_▲TT …5'
Bsa29 I	5'… AT^▼CGAT …3' 3'… TAGC_▲TA …5'	BanIII BseCI BspDI BspXI Bsu15I BsuTUI ClaI ZhoI	Isoschizomer	5'… AT^▼CGAT …3' 3'… TAGC_▲TA …5'
Bsc4 I	5'… CCNNNNN^▼NNGG …3' 3'… GGNN_▲NNNNNCC …5'	BseLI BsiYI BslI	Isoschizomer	5'… CCNNNNN^▼NNGG …3' 3'… GGNN_▲NNNNNCC …5'
Bse1 I	5'… ACTGGN^▼ …3' 3'… TGAC_▲CN …5'	BseNI BsrI BsrSI	Isoschizomer	5'… ACTGGN^▼ …3' 3'… TGAC_▲CN …5'
Bse3D I	5'… GCAATGNN^▼ …3' 3'… CGTTAC_▲NN …5'	BseMI BsrDI	Isoschizomer	5'… GCAATGNN^▼ …3' 3'… CGTTAC_▲NN …5'
Bse8 I	5'… GATNN^▼NNATC …3' 3'… CTANN_▲NNTAG …5'	BsaBI BseJI MamI	Isoschizomer	5'… GATNN^▼NNATC …3' 3'… CTANN_▲NNTAG …5'
Bse21 I	5'… CC^▼TNAGG …3' 3'… GGANT_▲CC …5'	AxyI Bsu36I Eco81I SauI	Isoschizomer	5'… CC^▼TNAGG …3' 3'… GGANT_▲CC …5'