METHOD FOR INSERTION OF NECESSARY DNA INTO MAMMAL CELL GENOME AND VECTOR SYSTEM FOR ITS REALIZATION Russian patent published in 2004 - IPC

FIELD: genetic engineering, molecular biology, biochemistry, biotechnological industry.

SUBSTANCE: invention proposes a method for integration of interesting DNA into the genome site of mammalian cell characterizing by the high transcription activity. Indicated site is marked preliminary by insertion of the specially constructed plasmid ("marker") into the cell and this plasmid comprises: a) DNA fragment that is heterologous to the cell genome after integration into genome and forms unique site for homologous recombination; b) DNA fragment encoding part of the first selective marker; c) at least one marker DNA sequence providing the possibility for cells selection wherein integration of "marker" plasmid has occurred with success. Cells selected from the first stage are transformed with the second ("target") plasmid comprising: a) DNA fragment eliciting the sufficient homology for carrying out the recombination with unique site(s) of "marker" plasmid; b) DNA fragment encoding the second part of the first selective marker that in the common expression with element (b) of the "marker" plasmid provides synthesis of the full marker protein; c) DNA that is suggested to be inserted into genome. By screening for expression of the first selective marker cells wherein DNA of the "target" plasmid and. Respectively, the "necessary" DNA are incorporated into genome DNA are selected. Invention proposes a set for realization of method comprising at least "marker" and "target" plasmids. The application of invention provides high level of expression in preparing any recombinant proteins.

EFFECT: improved insertion method, valuable biological properties of vectors system.

41 cl, 10 dwg, 5 tbl, 7 ex