Single Stage Purification of CRISPR/Cas13a Nuclease via Metal-Chelating Chromatography Following Heterologous Expression with the Preservation of Collateral Ribonuclease Activity

Autor:	N. D. Durmanov, O. I. Kiseleva, L. K. Kurbatov, A. V. Lisitsa, S. V. Kravchenko, S.P. Radko
Rok vydání:	2020
Předmět:	0106 biological sciences 0301 basic medicine Nuclease Chromatography biology 01 natural sciences Applied Microbiology and Biotechnology Biochemistry law.invention 03 medical and health sciences chemistry.chemical_compound 030104 developmental biology chemistry law 010608 biotechnology Recombinant DNA biology.protein CRISPR Chelation Heterologous expression Ribonuclease Polyhistidine-tag Biosensor
Zdroj:	Applied Biochemistry and Microbiology. 56:671-677
ISSN:	1608-3024 0003-6838
DOI:	10.1134/s0003683820060071
Popis:	CRISPR/Cas13a nucleases are currently considered to be the basis for the development of a new generation of biosensors for the ultrasensitive, in-field detection of bacterial and viral pathogens. A recombinant Cas13a nuclease with functional affinity was obtained as a result of heterologous expression in E. coli with a single-step purification process via metal-chelating chromatography with the N-terminal polyhistidine tag. The simplified procedure of Cas13a nuclease purification broadens the possibilities for the development and practical application of diagnostic biosensing systems based on it. Moreover, our results indicate that the currently uncharacterized protein U2PWF1 of Leptotrichia wadei represents Cas13a nuclease.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_________::6c6cc8653b4ad9387aff7cfbf55ef220 https://doi.org/10.1134/s0003683820060071 Zobrazit plný text záznamu Plný text ve formátu PDF Plný text ve formátu HTML
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