Journal article
Authors list: Ulyanova, V; Mahmud, RS; Klug, G; Preissner, KT; Vershinina, V; Ilinskaya, O
Publication year: 2017
Pages: 222-225
Journal: BioNanoScience
Volume number: 7
Issue number: 1
DOI Link: https://doi.org/10.1007/s12668-016-0349-z
Publisher: Springer
Low molecular weight guanyl-preferring ribonuclease (RNase) secreted by Bacillus pumilus
Abstract:
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.
Citation Styles
Harvard Citation style: Ulyanova, 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 style: Ulyanova, 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