MODIFICATION OF THE YEAST-2-HYBRID SYSTEM MAKING IT POSSIBLE TO TEST INTERACTIONS BETWEEN PROTEINS WITHIN MULTICOMPONENT PROTEIN COMPLEXES Russian patent published in 2021 - IPC C12N1/19 C12N15/81 C12P21/00 

FIELD: biotechnology.

SUBSTANCE: invention relates to a yeast cell designed to detect interactions between proteins and their domains in multicomponent protein complexes. The yeast cell is co-transformed by three plasmids modified to express several proteins in yeast cells. In this case, the nucleotide sequence encoding the DNA-binding domain of the GAL4 protein is fused with the nucleotide sequence encoding the first protein and cloned into one plasmid. The nucleotide sequence encoding the activation domain of the GAL4 protein is fused with a nucleotide sequence encoding the second and third proteins and having a single reading frame with one promoter and terminator, and cloned into a second plasmid, while a nucleotide sequence encoding a self-cleaving peptide is embedded between the second and third proteins, which makes it possible for several proteins or their domains to be expressed simultaneously in a single reading frame with one promoter and terminator. The nucleotide sequence encoding the non-hybrid protein has been cloned into a third plasmid. In this case, the self-cleaving peptide is a 2A-peptide of the horse B rhinitis virus, having the amino acid sequence GlySerGlyGlyAlaThrAsnPheSerLeuLeuLysLeuAlaGlyAspValGluLeuAsnProGlyPro, presented in SEQ ID NO: 1.

EFFECT: invention can be used to detect interactions between protein molecules as part of multicomponent protein complexes that can be used for medical and research purposes.

1 cl, 2 dwg, 2 ex