RECOMBINANT PLASMID DNA pcDNA4-Apo-2NLS2 CARRYING SYNTHETIC GENE OF APOPTIN PROTEIN INDUCING p53-INDEPENDENT PROGRAMMED TUMOUR CELL DEATH Russian patent published in 2015 - IPC C12N15/34 

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to molecular biology, more specifically to recombinant expression of apoptin, and can be used as an anti-tumour therapeutic transgene. The engineered plasmid DNA pcDNA4-Apo-2NLS2 has a molecular weight of 5.13 mD, a size of 7,896 base pairs and contains in accordance with a physical and genetic map presented in Fig. 5, an operon having a size of 1,071 base pairs consisting of the early late promoter CMV, a leader fragment, a sequence of the penetrating peptide of TAT HIV protein, an apoptin gene of chick anaemia virus and a sequence of Flag epitope; the promoter CMV of 232-819 base pairs; T7 promoter of 863-882 base pairs; Xpres primer of 1,112-1,130 base pairs; TOPO cloning site of 1,184 base pairs; genetic markers: the resistance gene Zeocin; b-lactamase gene determining a resistance to ampicillin; unique recognition sites: ClaI (1918); XbaI (2619); BamHI (2993); SalI (3001); BstWI (3425). The produced plasmid DNA pcDNA4-Apo-2NLS2 applied for the apoptin protein expression with the double site NLS2 selectively inducing the p53-independent programmed death of individual's neoplastic tumour cells. The synthetic apoptin gene as a component of the above DNA is characterised by a nucleotide sequence SEQ ID NO1 and a size of 393 base pairs.

EFFECT: invention enables providing higher activity of the expressed modified apoptin protein gene in relation to inducing the selective programmed tumour cell death.

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