pLV-SFFV-hTYR-PGK-Puro
- 20 ug / Standard ((DNA1057-STAN)) $ 1,500
- Name of plasmid: pLV-SFFV-hTYR-PGK-Puro
- Description: pHR' SIN-SFFV-hTYR-PGK-Puro plasmid DNA
- Amount of DNA: 20 ug
- Promoter: SFFV: spleen focus forming virus promoter for high constitutive gene expression in numerous cell types. Unlike the CMV promoter, SFFV works very well in human, rodent, and other animal cells. PGK: mouse phosphoglycerate kinase I promoter drives puromycin resistance for selection of transduced cells.
- Transgenes: Human tyrosinase (hTYR) and puromycin resistance (Puro)
- Proposed Use: For generation of lentiviral vector stocks encoding human tyrosinase and puromycin resistance in a triple transfection protocol.
Plasmid Map:
Transgene Validation: Cells were transduced with LV-hTYR-PGK-Puro, selected with puromycin, and lysed. The lysates were incubated with L-DOPA, which in the presence of tyrosinase is converted to a colored compound with absorbance at 475 nm. B16F10 cells (endogenously express tyrosinase) and parental cells were used as controls.