Why does Lonza recommend constructing a double gene vector (DGV) before transfection? |
 | The generation of stable cell lines following transfection of mammalian cells is the result of random integration of DNA into the genome of the host cell.
The simplest way to reduce any disruption of the heavy and light chain genes is to join them together before transfection. After transfection, the most likely scenario is that there is an equal number of each gene integrated into a single locus in the genome. The weak promoter on the GS gene enables selection of cell lines where the construct has integrated at transcriptionally active regions of the genome.
Use of a double-gene vector facilitates the integration of the product genes at the same transcriptionally active site, which enables the selection of high-producing cell lines using the GS System™. |
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