IN VITRO METHODS FOR INDUCTION AND MAINTENANCE OF PLANT CELL LINES AS SINGLE SUSPENSION CELLS WITH INTACT CELL WALLS AND TRANSFORMATION THEREOF Russian patent published in 2012 - IPC C12N5/00 

FIELD: chemistry.

SUBSTANCE: invention relates to biotechnology. Described is a method of obtaining a separate plant cell, where the method involves: putting aggregated plant cells having intact cell walls into a medium containing glycerol and a separating agent selected from a group consisting of a pectin-degrading enzyme, and a tubulin de-polymerising compound; shaking the aggregated plant cells from step (a) at temperature 25-28°C in the dark on an orbital shaker at 150 rpm; and transferring one or more plant cells, having intact walls, into the plant culturing medium which does not contain glycerol and a separating agent, selected from a group consisting of a pectin-degrading enzyme and a tubulin de-polymerising compound. Disclosed is a method of transforming a cell obtained using the described method, wherein said method involves obtaining an isolated cell according to the described method, transformation of said cell with a heterologous polynucleotide and collecting the transformed cell. The invention simplifies and integrates transformation methods based on separate cells.

EFFECT: high-efficient production of marker-free and uniform transgenic lines for meeting different needs when implementing programs associated with animal health, biopharmaceutics and crop protection.