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Next Generation DNase for Ultimate Performance in Molecular Biology Applications
EP31093
Poster Title: Next Generation DNase for Ultimate Performance in Molecular Biology Applications
Submitted on 16 Dec 2019
Author(s): Darius Kavaliauskas , Viktorija Vitkovskė , Gediminas Alzbutas , Juozas Šiurkus
Affiliations: Thermo Fisher Scientific Baltics
Poster Views: 536
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Poster Information
Abstract: We have developed a new recombinant DNase I which is
produced by GRAS recombinant host for mRNA synthesis
workflow, by taking into account all quality attributes which are
required for such enzyme and application. In addition the new DNAse I mutant was engineered which is able to hydrolyze DNA at higher ionic strength and features increased thermolability if compared to conventional recombinant DNAse I. This next generation DNAse mutant
could be an alternative for DNase I in various mRNA workflow
applications where additional features of higher salt tolerance
and thermolability are important. In our work we have compared the key enzymatic features of these enzymes and demonstrated various application scenarios which are common for different mRNA workflows.
Summary: Salt tolerance studies showed that the new DNAse I
mutant is much more salt tolerant than wild type DNAse I
and has a comparable salt tolerance as Turbo DNase. Thermolability experiments revealed that DNAse I mutant is more thermally labile than wild type DNAse I and slightly more labile than Turbo DNase. DNase I mutant hydrolyzes single and double stranded DNA substrates at a rate similar to wild type DNase I.
References: 1. Hyone Myong Eun , in Enzymology Primer for Recombinant
DNA Technology, 1996.
2. Surya Pandey Taro Kawai, in Biological DNA Sensor, 2014.
3. Tatiana Borodina , Marc Sultan et al., in Methods in
Enzymology ,
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