METHOD OF HOMINAL MITOCHONDRIAL GENOME COPYING IN YARROWIA LIPOLYTICA YEAST CELLS Russian patent published in 2018 - IPC C12N15/63 C12N15/81 C12N1/19 

FIELD: biotechnology.

SUBSTANCE: method of hominal mitochondrial genome copying in yarrowia lipolytica yeast cells is proposed. Using the method of chemical transformation, the preparation of the total human DNA is transformed into a Yarrowia lipolytica strain containing the previously integrated pQ-SRUS and Lys39H (SEQ ID NO 1) constructs based on the hygromycin resistance marker HygR (SEQ ID NO 3) or the Yarrowia lipolytica strain, containing previously integrated constructs of pQ-SRUS and Leu36H (SEQ ID NO 2). Transformants are selected on a minimal synthetic medium containing succinate as the sole energy source. The presence of replicating DNA of the human mitochondrial genome is determined using real-time PCR with pairs of primers specific for the DNA of the mitochondrial human genome: MTTL1 (gggtttgttAagatggcagagcccggt)/MTTL2 (ggttggccatgggtatgttgttaagaa) to identify the sequence of the human tRNA gene Leu genome and TRNK1 (agaaccaacacctctttacagtgaaat)/TRNK2 (ttagttgggtgatgaggaatagtgtaa) to identify the sequence of the human TPHK-Lys gene.

EFFECT: invention can be used to correct harmful mutations in the mitochondrial human genome and the preparation work corrected for the sequence of intact mitochondrial DNA in yeast cells for the purpose of its subsequent introduction into mesenchymal stromal cells of the patient for the treatment of widespread mitochondrial human diseases that violate the functionality of the mitochondrial Lys-Leu-specific ribosomal riziform.

6 dwg, 1 tbl