METHOD FOR SPECIFIC SELECTION OF DNA MOLECULES (DNA APTAMERS) WITH AFFINITY FOR RECOMBINANT PROTEIN TARGET Russian patent published in 2014 - IPC C12N15/63 C07H21/04 C12N15/66 C12N15/72 A61K31/7088 G01N33/02 G01N33/15 G01N33/48 

Abstract RU 2513700 C1

FIELD: chemistry.

SUBSTANCE: invention relates to biochemistry, particularly a method for specific collection of DNA molecules (DNA aptamers) with high affinity for a recombinant protein target. Said method involves synthesis of a single polypeptide chain of a recombinant protein containing a fragment of glutathione S-transferase, a protein target, a peptide sequence split by the B. Anthracis lethal factor, a peptide which is biotinylated in vitro under the action of an E.coli biotin-ligase enzyme, binding the obtained recombinant polypeptide with an oligonucleotide library and immobilising the protein on paramagnetic particles bearing glutathione, washing the paramagnetic particles with the immobilised polypeptide from unbound oligonucleotides in a liquid stream, splitting the protein target with the bound DNA aptamers from the surface of paramagnetic particles with the B. anthracis lethal factor, separating and amplifying the DNA sequence with affinity to the recombinant protein target in a polymerase chain reaction and obtaining a set of single-chain DNA aptamers that are specific to the protein target.

EFFECT: invention provides efficient production of DNA aptamers with high affinity for recombinant protein targets.

4 dwg, 4 ex

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RU 2 513 700 C1

Authors

Kozyr' Arina Vladimirovna

Kolesnikov Aleksandr Vladimirovich

Luneva Nina Mikhajlovna

Khlyntseva Anna Evgen'Evna

Shemjakin Igor' Georgievich

Dates

2014-04-20Published

2012-09-27Filed