Cell transfection is a technique for introducing exogenous molecules (e.g., DNA, RNA, etc.) into eukaryotic cells.
Transfection Classification: Transient transfection and stable transfection.
Transient Transfection: The exogenous DNA or RNA is not integrated into the host chromosome for the expression of exogenous target genes after being transferred into the host cell.
Stable Transfection: The exogenous target gene is integrated into the host chromosome for expression after the exogenous DNA is transferred into the host cell.
1. Integration of the target gene into the chromosome requires the use of linear DNA (linear DNA has a higher integration rate although the amount of DNA transferred is lower than that of superhelical DNA)
2. The integration of exogenous DNA into the chromosome of the host cell is about 1%, and it is usually necessary to repeatedly screen by some selective markers to obtain a stable transfection of homologous cell lines.
3. Replicates, transcribes, translates, and is stably inherited in the host cell genome.
4. Single-cell clones used to obtain stably expressed exogenous genes: e.g. large-scale protein synthesis, long-term pharmacological studies of genetic mechanisms, etc.
Stabilized Cell Line: Generally, the exogenous DNA is cloned into a vector with a certain resistance or selection gene, the vector is transfected into the host cell and integrated into the chromosome, and the selection marker contained in the vector is used for screening to obtain a cell line that can stably express the target protein.
Stable Cell Line Construction Process
High Quality: Exogenous genes can be strictly identified, clear cell source and free of contamination.
High Yield: Recombinant protein/antibody expression can be rapidly optimized to above 2g/L.
Stable: The unique GS stable transfection series of vectors, together with MSX drug screening, realize stable transmission for more than 15 generations.
Rapid: High yielding cell lines can be screened in as little as 3 months.
Customized: Personalized expression of recombinant proteins/antibodies in full-length or fragmented form
Experienced: Professional technical team, delivered stable cell lines for many domestic and foreign pharmaceutical companies and research units.
Multi-screening System: G418/tideomycin/puromycin screening system, GS/DHFR dual gene screening system, etc.
Stable Cell Line Construction
Gene Sequences Vector
Recombinant expression plasmid construction
1-3 Cell lines
Technical service reports
Determination of drug background concentration
Screening and identification of polyclonal stable cell lines
Screening and identification of monoclonal stable cell line
Stabilized cell line yield, activity test and stability study(optional)
Construction of CHO stabilized cell lines for the production of recombinant antibodies