Journal article

A Set of Genetic Constructs for Binase and Barstar Overproduction


Authors listUlyanova, V; Mahmud, RS; Klug, G; Preissner, KT; Vershinina, V; Ilinskaya, O

Publication year2017

Pages222-225

JournalBioNanoScience

Volume number7

Issue number1

DOI Linkhttps://doi.org/10.1007/s12668-016-0349-z

PublisherSpringer


Abstract

Low molecular weight guanyl-preferring ribonuclease (RNase) secreted by Bacillus pumilus
is referred to as binase and regarded as a potential agent to target
oncogenic diseases and viral infections. Depending on research purposes,
native binase or tagged and mutated forms are required for application.
Specific inhibition of binase activity is achieved by binding to RNase
inhibitor barstar. Native and mutant forms of binase are routinely
obtained by ion-exchange chromatography, while tagged versions of
binase, which can be purified faster and less tedious than non-tagged,
are currently unavailable. Here, recombinant DNA techniques were
employed to improve binase overexpression in bacterial host cells, its
purification, and subsequent detection in downstream assays using
affinity tag. Here, recombinant DNA techniques were employed to improve
binase overexpression in bacterial host cells. Binase-barstar genetic
cassette was modified to obtain strong two-gene operon with the
possibility of rapid exchange of binase and/or barstar genes with
homologous or mutated ones as well as the generation of C-terminally
His-tagged binase and/or barstar. The binase-barstar operon was
introduced into pET26b vector as well as into pET26b-pET15b hybrid
vector that was created in this study. As a result, plasmids allowed the
induced production of extracellular non-tagged, N- or C-terminal
His-tagged binase and synchronous intracellular production of non-tagged
or C-His-tagged barstar. The constructs can be further sub-cloned in
gram-positive bacterial hosts to ensure native production conditions.




Authors/Editors




Citation Styles

Harvard Citation styleUlyanova, V., Mahmud, R., Klug, G., Preissner, K., Vershinina, V. and Ilinskaya, O. (2017) A Set of Genetic Constructs for Binase and Barstar Overproduction, BioNanoScience, 7(1), pp. 222-225. https://doi.org/10.1007/s12668-016-0349-z

APA Citation styleUlyanova, V., Mahmud, R., Klug, G., Preissner, K., Vershinina, V., & Ilinskaya, O. (2017). A Set of Genetic Constructs for Binase and Barstar Overproduction. BioNanoScience. 7(1), 222-225. https://doi.org/10.1007/s12668-016-0349-z


Last updated on 2025-21-05 at 15:11