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Alteration of sequence specificity of the type IIS restriction endonuclease BtsI.

التفاصيل البيبلوغرافية
العنوان: Alteration of sequence specificity of the type IIS restriction endonuclease BtsI.
المؤلفون: Shengxi Guan, Aine Blanchard, Penghua Zhang, Zhenyu Zhu
المصدر: PLoS ONE, Vol 5, Iss 7, p e11787 (2010)
بيانات النشر: Public Library of Science (PLoS), 2010.
سنة النشر: 2010
المجموعة: LCC:Medicine
LCC:Science
مصطلحات موضوعية: Medicine, Science
الوصف: The Type IIS restriction endonuclease BtsI recognizes and digests at GCAGTG(2/0). It comprises two subunits: BtsIA and BtsIB. The BtsIB subunit contains the recognition domain, one catalytic domain for bottom strand nicking and part of the catalytic domain for the top strand nicking. BtsIA has the rest of the catalytic domain that is responsible for the DNA top strand nicking. BtsIA alone has no activity unless it mixes with BtsIB to reconstitute the BtsI activity. During characterization of the enzyme, we identified a BtsIB mutant R119A found to have a different digestion pattern from the wild type BtsI. After characterization, we found that BtsIB(R119A) is a novel restriction enzyme with a previously unreported recognition sequence CAGTG(2/0), which is named as BtsI-1. Compared with wild type BtsI, BtsI-1 showed different relative activities in NEB restriction enzyme reaction buffers NEB1, NEB2, NEB3 and NEB4 and less star activity. Similar to the wild type BtsIB subunit, the BtsI-1 B subunit alone can act as a bottom nicking enzyme recognizing CAGTG(-/0). This is the first successful case of a specificity change among this restriction endonuclease type.
نوع الوثيقة: article
وصف الملف: electronic resource
اللغة: English
تدمد: 1932-6203
Relation: http://europepmc.org/articles/PMC2909958?pdf=render; https://doaj.org/toc/1932-6203
DOI: 10.1371/journal.pone.0011787
URL الوصول: https://doaj.org/article/de23d2bea7b54095ba863f00187505bf
رقم الأكسشن: edsdoj.23d2bea7b54095ba863f00187505bf
قاعدة البيانات: Directory of Open Access Journals
الوصف
تدمد:19326203
DOI:10.1371/journal.pone.0011787