Seminar of Cell Culture Techniques. Tapodi Antal Department of Biochemistry and Medicinal Chemistry, Faculty of Medicine, University of Pecs, Hungary . I . Cells Types II . Introduction to Cell Culture Lab III . Techniques. Content s. Prim a r y cultures Secondary cultures Normal
Department of Biochemistry and Medicinal Chemistry, Faculty of Medicine, University of Pecs, Hungary
HGPRT and TK-/- myeloma and B-cells from immunized animal
Metabolic pathways relevant to hybrid selection in medium containing hypoxanthine, aminopterin andthymidine (HAT medium).
When the main synthetic pathways are blocked with the folic acid analogue aminopterin (*), the cell must depend on the “salvage” enzymes HGPRT and TK (thymidine kinase). HGPRT (-) cells cannot grow in HAT medium unless they are fused with HGPRT (+) cells.
Hypoxanthine Inosine Monophosphate
Guanine Guanosine Monophosphate
Thymidine GDP dGDP
Thymidine kinase RNA GTP dGTP
dTMP dTDP d TTP DNA
UDP dUTP dUMP dCTP dATP
Reaction catalyzed by PARP
pEGFP with NLS
pEGFP without NLS
two copies of chromosome 3 (red), four copies of chromosome 7 (green), five copies of chromosome 17 (aqua) and one copy of p16 gene (gold)