The Type IIS restriction enzyme Eco47I, also known as R.Eco47I, is a prokaryotic DNA methyltransferase that is involved in the detection of CG-specific methylation [1]. This enzyme is a thermostable restriction endonuclease extracted from extreme thermophilic bacteria, and it is an isoschizomer of prototype Type II restriction endonucleases AvaII and BstYI [2]. The use of type IIS restriction enzymes, such as Eco47I, presents a unique strategy for efficient cloning in protein crystallography [3]. Additionally, the involvement of Type II restriction endonucleases, including Eco47I, in biological restriction has been demonstrated, indicating their significance in DNA recognition specificity [4]. Furthermore, type IIS restriction enzymes, like Eco47I, are utilized in hierarchical DNA assembly systems due to their ability to block restriction sites and generate sticky ends for efficient DNA piece joining [5]. Moreover, type IIS restriction enzymes, which recognize sequences outside the cleavage sites, allow a programmable signature and can be used in an alternating pattern to form a ‘cloning loop’ within a proprietary vector [6][7].
References:
[1] P. Albert, B. Varga, N. Zsibrita, & A. Kiss, "Circularly permuted variants of two cg-specific prokaryotic dna methyltransferases", Plos One, vol. 13, no. 5, p. e0197232, 2018. https://doi.org/10.1371/journal.pone.0197232
[2] D. Gupta and N. Sharma, "Thermostable restriction endonucleases from thermophilic bacteria", International Research Journal of Pharmacy, vol. 5, no. 4, p. 259-263, 2014. https://doi.org/10.7897/2230-8407.050455
[3] N. Galloway, H. Toutkoushian, M. Nune, N. Bose, & C. Momany, "Rapid cloning for protein crystallography using type iis restriction enzymes", Crystal Growth & Design, vol. 13, no. 7, p. 2833-2839, 2013. https://doi.org/10.1021/cg400171z
[4] P. Whitehead, D. Jacobs, & N. Brown, "Restriction endonucleases fromherpetosiphon giganteus: an example of the evolution of dna recognition specificity?", Nucleic Acids Research, vol. 14, no. 17, p. 7031-7045, 1986. https://doi.org/10.1093/nar/14.17.7031
[5] L. Da and C. O’Callaghan, "Metclo: methylase-assisted hierarchical dna assembly using a single type iis restriction enzyme", Nucleic Acids Research, 2018. https://doi.org/10.1093/nar/gky596
[6] A. Sarrión-Perdigones, M. Vázquez‐Vilar, J. Palací, B. Castelijns, J. Forment, P. Ziarsoloet al., "Goldenbraid 2.0: a comprehensive dna assembly framework for plant synthetic biology", Plant Physiology, vol. 162, no. 3, p. 1618-1631, 2013. https://doi.org/10.1104/pp.113.217661
[7] H. DePaoli, G. Tuskan, & X. Yang, "An innovative platform for quick and flexible joining of assorted dna fragments", Scientific Reports, vol. 6, no. 1, 2016. https://doi.org/10.1038/srep19278
