KasI vs. SspDI: Comparative analysis of enzyme activity for restriction digestion

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dc.contributor.authorSumanen_US
dc.contributor.authorPuria, Rekhaen_US
dc.contributor.authorNain, Vikranten_US
dc.date.accessioned2020-01-02T06:38:38Z
dc.date.available2020-01-02T06:38:38Z
dc.date.issued2019-12
dc.description.abstractType II restriction enzymes are routinely used by molecular biologists in designing and implementation of cloning experiments without referring to the literature on enzymes in use, and at times, face some unforeseeable problems. In our laboratory too, we encountered one such problem while working with KasI restriction enzyme which recognizes GGCGCC sequence, and we further analyzed the issue. Our observations corroborate the fact that KasI acts as monomer and cleaves double stranded DNA through nicking mechanism. It introduces breaks in two strands of DNA after substantial time gap which can be owed to two independent nickase activities in the opposite strands. Moreover, this time gap between two nickase activities results in formation of different topological forms of DNA. Since molecular biologists working with common restriction enzymes are not familiar with such nickase activity, they may misinterpret their restriction digestion results. However, no such problem was observed with the use of SspDI restriction enzyme which also recognizes the same sequence (GGCGCC) and produces the identical overhangs as by KasI. Hence, SspDI suits better for routine cloning and genetic modification purposes over KasI while using GGCGCC as cloning site.en_US
dc.identifier.affiliationsSchool of Biotechnology, Gautam Buddha University, Greater Noida-201 312, Uttar Pradesh, Indiaen_US
dc.identifier.citationSuman, Puria Rekha, Nain Vikrant. Individual and combined exposure of cupric chloride di-hydrate and anthracene to Lamellidens corrianus (Lea): Evaluation of genotoxic responses using comet assay. Indian Journal of Experimental Biology. 2019 Dec; 57(12): 949-955en_US
dc.identifier.issn0975-1009
dc.identifier.issn0019-5189
dc.identifier.placeIndiaen_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/191426
dc.languageenen_US
dc.publisherNISCAIR-CSIR, Indiaen_US
dc.relation.issuenumber12en_US
dc.relation.volume57en_US
dc.source.urihttps://nopr.niscair.res.in/handle/123456789/52199en_US
dc.titleKasI vs. SspDI: Comparative analysis of enzyme activity for restriction digestionen_US
dc.typeJournal Articleen_US
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