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Amino Acid Sequence: Variation of mVENUS, with L69V and K80R mutations
Applications: Optimizing transduction assays
Easily track transduction efficiency over time
Generation of YFP (Topaz) Reporter stable cell lines
Description: Topaz1, a yellow fluorescence protein (YFP), is one of the brightest and thermally stable green fluorescence proteins (GFP). The YFP (Topaz) Reporter Lentivirus contains replication-defective, VSV-G pseudotyped lentiviral particles that are ready to be transduced into almost all types of mammalian cells, including primary and non-dividing cells. The particles contain an EF1α promoter-driven YFP(Topaz) construct (a.a. sequence below), and confer both YFP (Topaz) expression and puromycin resistance to the target cells. YFP (Topaz) expression and transduction efficiency can easily be verified and optimized via fluorescence microscopy or flow cytometry. Topaz has an excitation wavelength of 514 nm, an emission wavelength of 527 nm, and extinction coefficient of 94,500M-1cm-1.
Genbank: AUG68261
Storage Stability: Lentiviruses are shipped with dry ice. For long term storage, it is recommended to store the virus at -80°C. Avoid repeated freeze-thaw cycles. Titers can drop significantly with each freeze-thaw cycle.
Supplied As: Two vials (2 x 500 µl) of YFP (Topaz) reporter lentivirus at a titer ≥5 x 10^6 TU/ml. The titer will vary with each lot, and multiplicity of infection will depend on the target cell type. We provide a standardized p24 measurement with each shipment
Warnings: Biosafety: The lentiviruses are produced with the third generation SIN (self-inactivation) lentivector which ensures self-inactivation of the lentiviral construct after transduction and integration into the genomic DNA of the target cells. None of the HIV genes (gag, pol, rev) will be expressed in the transduced cells, as they are expressed from packaging plasmids lacking the packing signal. Although the pseudotyped lentiviruses are replication-incompetent, they require the use of a Biosafety Level 2 facility. BPS recommends following all local federal, state, and institutional regulations and using all appropriate safety precautions.
Biosafety Level: BSL-2
References: Cubitt, A.B. et. al., Understanding structure-function relationship in the Aequorea victoria green fluorescence protein, Methods in Cell Biology, 1998;58:19-30.