FIELD: biotechnology.
SUBSTANCE: invention relates to the field of molecular genetics and medical biotechnology and can be used in medicine for designing technology of differential diagnosis of autoimmune diabetes, stiff-person syndrome and other diseases. A method has been developed for transfection and cultivation of cells synthesizing recombinant protein - glutamate decarboxylase GAD65, based on transfection of HEK293 cell culture with a specially constructed plasmid containing human protein GAD65 gene under the eukaryotic promotor. Stable gene expression and synthesis of GAD65 protein in transfected HEK293 cells is achieved through regular additional enrichment of the culture medium with glutamine.
EFFECT: proposed method for cultivation of GAD65-producer cells in a medium with increased glutamine content allows ensuring stable synthesis of glutamate decarboxylase under laboratory conditions with simultaneous increased transfection efficiency.
1 cl, 2 dwg, 3 tbl, 3 ex
| Title | Year | Author | Number |
|---|---|---|---|
| GENETIC NUCLEOTIDE SEQUENCES OF ARTIFICIALLY MODIFIED GDNF WITH DELETED pro-REGION, PRODUCT OF WHICH HAS IMPROVED PROPERTIES NEURAL INDUCTOR AND STIMULATOR OF FORMATION OF NEURAL PROCESSES, SUITABLE FOR THERAPY OF NEURAL INJURIES, ISCHEMIC STROKES AND NEURODEGENERATIVE DISEASES, EXPRESSION VECTOR, MODIFIED WITH GDNF | 2014 |
|
RU2595377C2 |
| METHOD OF OBTAINING RECOMBINANT VESICULAR STOMATITIS VIRUS | 2022 |
|
RU2807751C1 |
| MONOMOLECULAR CHIMERIC T-CELLULAR RECEPTOR TO CANCER-EMBRYONIC ANTIGEN | 2015 |
|
RU2652955C1 |
| MONOMOLECULAR CHIMERIC T-CELL RECEPTOR TO CANCER ANTIGEN CA125 | 2018 |
|
RU2747095C2 |
| METHODS AND FORMULATIONS FOR CELLS TRANSFECTION | 2012 |
|
RU2624139C2 |
| METHOD OF PRODUCING TARGET VIRUS OR TARGET PROTEINS DIFFERENT FROM ADENOVIRUS, EXPRESSION CELL AND HOST CELL AND PRODUCTION METHODS THEREOF, USE OF EXPRESSION CELL | 2006 |
|
RU2449015C2 |
| METHODS AND PREPARATIONS FOR TRANSFECTION OF CELLS | 2012 |
|
RU2691027C2 |
| METHOD FOR PRODUCING TRANSGENIC BIRDS USING EMBRYO STEM CELLS | 2007 |
|
RU2473688C2 |
| METHOD FOR PRODUCING REPLICATIVE INFLUENZA VIRUS PARTICLES, CELL COMPOSITION (VERSIONS), CELL CULTURE COMPOSITION AND APPLICATION THEREOF | 2005 |
|
RU2435855C2 |
| CULTURE CELL STRAIN CHO-IL7/13 PRODUCING HUMAN INTERLEUKIN-7 | 2012 |
|
RU2562169C2 |
