Fluorescent Protein Vectors
|The Vivid Colors™ Fluorescent Protein Vectors are Invitrogen' next-generation cloning and expression tools featuring Aequorea victoria-derived fluorescent proteins (FP) for simple, non-invasive detection of recombinant proteins. With Gateway® and TOPO® cloning vector options; cloning, expression and viewing has never been so easy! |
Spectral Properties of Vivid Colors™ Fluorescent Proteins
Expression of the Vivid Colors™ EmGFP and YFP fusion proteins can be detected by fluorescence microscopy or flow cytometry using standard FITC filter sets. For optimal visibility, it is recommended that you use a filter set optimized for detection within the excitation and emission ranges for each fluorescent protein (see table below). Excitation and emission profiles for all FPs are also illustrated.
|EmGFP||Green||487||509||S65T, S72A, N149K, M153T, I167T||XF100|| |
|YFP||Yellow||514||527||S65G, S72A, K79R, T203Y||XF1042||41028|
|CFP||Cyan||452||505||K26R, Y66W, N146I, M153T, V163A, N164H||XF114||31044|
|BFP||Blue||308-383||440-447||F64L, Y66H, Y145F, V163A , N198S||XF10||31021|
|Cycle 3 GFP||Green||395478||507||F64L, F100S, M154T, V164A||XF76|| |
Vivid Colors™ Fluorescent Protein Fusion Vectors
These Vivid Colors™ vectors are for generation of N-terminal or C-terminal Fluorescent Protein fusions to your Gene-Of-Interest (GOI). Emerald Green (EmGFP) and Yellow (YFP) Fluorescent Protein versions are available. To create a Vivid Colors™ fusion with protein, you can employ either Gateway® or TOPO®-cloning technology. Gateway® Destination Vectors require the use of an entry clone, while the TOPO® vectors require only a PCR product of your GOI (T Cloning®). Regardless of the cloning techniques you select, the Ultimate ORF™ collection is a comprehensive and convenient source of full-length cDNA clones for creating fusion proteins.
Co-expression of Your Protein with a Vivid Colors™ Fluorescent Protein
Express your protein in it’s native-protein configuration AND EmGFP from a single vector. The Vivid Colors™ pcDNA™ 6.2/ EmGFP-Bsd / V5-DEST vector allows you to rapidly identify transfected cells to look for biological activity conferred by your gene. You’ll need to use a Gateway® entry-clone version of your gene to clone into this destination (DEST) vector. The Ultimate ORF™ collection is a comprehensive and convenient source of full-length entry clones for this purpose. If you already have your a clone of your protein, you can amplify it by PCR and clone it with TOPO®-technology to prepare the entry-clone.
Create Your Own Vectors with Other Vivid Colors™ Fluoresecent Proteins
The pRSET/CFP (cyan) and pRSET/BFP (blue) vectors contain alternative color versions of fluorescent proteins derived from Aequorea victoria. These proteins are commonly used with the YFP and EmGFP for multi-color applications. Configured for bacterial expression and fluorescent protein purification, these vectors can also be used for subcloning to construct customized mammalian expression vectors.
Lentiviral (Vivid Color™) Emerald GFP Control Expression Vector
To optimize lentivial packaging and transduction efficiencies and monitor infected cells, you need the pLenti 6.2/EmGFP-GW Expression Control Vector. This vector allows you to generate high-titer Lentiviral preps that express EmGFP and provides for blasticidin selection. To learn more about the Lentiviral Expression product line visit our ViraPower™ catalog pages.
Other Fluorescent Protein Vectors (Cycle 3 Green Fluorescent Protein)
To express your gene with alternative promoters, or with other selectable markers for stable-cell generation, consider the pTracer vectors series. These vectors express the Cycle 3 Green Fluorescent Protein (Cycle 3 GFP) which has spectral excitation and emission wavelengthsidentical to wild-type GFP. Although nearly as bright as the Vivid Colors™ FPs, you’ll need to use the proper filter set to observe the fully-enhanced brightness.
- pTracer™/Bsd Expression Vectors
Choose from CMV or EF-1 a promoters for expressing your gene of interest. The pTracer™ EF vector will allow expression of your protein with carboxy-terminal V-5 and 6xHis epitope tags. The pTracer™ CMV vector is for expression of your protein without any epitope tags. These pTracer™ /Bsd Vectors express the blasticidin resistance gene as a fusion product with the Cycle 3 GFP.
- pTracer™/Zeo Expression Vectors
Choose from SV40, CMV or EF-1 a promoters for expressing your gene of interest. The pTracer™ EF vector will allow expression of your protein with carboxy-terminal V-5 and 6xHis epitope tags. The pTracer™ SV40 and CMV vectors are for expression of your protein without any epitope tags. These pTracer™ /Bsd Vectors express the zeocin resistance gene as a fusion product with the Cycle 3 GFP.
- Promoter Characterization
If you’re trying to characterize an unknown promoter, you’ll need a cloning and reporter system to monitor expression efficiencies from your PCR-generated mutants. The pGlow and pBlue TOPO® TA Expression Kits provide the convenience of TOPO®-cloning of your PCR products, and the option to quantify promoter activity with either Cycle 3 GFP (pGlow) or b - galactosidase (pBlue) expression .
- GFP (cycle 3) TOPO® TA Expression Kits
These vectors are for cloning your PCR-amplified gene of interest to generate a fusion protein with Cycle 3 GFP fused on either the N- or C-termini. With TOPO®-cloning, only a 5-minute reaction is required to generate a clone. Following verification of orientation, the fusion protein is ready to express. The vector elements for these TOPO®-cloning vectors include a CMV promoter for high-level expression of your gene of interest and a neomycin resistance marker.
Highly Specific Anti-GFP Antibodies for Vivid Colors™ Fluorescent Proteins
Highly specific antibodies to Aequorea victoria fluorescent proteins are available, including polyclonal Anti-GFP (rabbit serum) and Anti-GFP (rabbit IgG fraction), for Western Blot detection. These antibodies have been tested and are shown to react specifically and strongly to both Vivid Colors™ EmGFP and YFP. A number of different conjugated forms of these antibodies are also available from Molecular Probes, offering you a choice of multiple detection technologies you can employ.
Anti-GFP; Rabbit polyclonal antiserum (A6455)
Anti-GFP; Rabbit polyclonal IgG fraction (A11122)
Anti-GFP; ABfinity™ recombinant rabbit monoclonal antibody (G10362)
Table 1 - Summary of Fluorescent Protein Vector Elements
|Gene of Interest||Mammalian|
|Vivid Colors™ pcDNA™ 6.2/C-EmGFP-DEST||Emerald GFP||C-terminal FP fusion||Gateway®||CMV|| ||bsd|
|Vivid Colors™ pcDNA™ 6.2/N-EmGFP-DEST||Emerald GFP||N-terminal FP fusion||Gateway®||CMV||N-term.V5||bsd|
|Vivid Colors™ pcDNA™ 6.2/C-YFP-DEST||Yellow||C-terminal FP fusion||Gateway®||CMV|| ||bsd|
|Vivid Colors™ pcDNA™ 6.2/N-YFP-DEST||Yellow||N-terminal FP fusion||Gateway®||CMV||N-term.V5||bsd|
|Vivid Colors™ pcDNA™ 6.2/ EmGFP-Bsd/V5-DEST||Emerald GFP||native||Gateway®||CMV|| ||GFP-bsd fusion|
|Vivid Colors™ pcDNA™ 6.2/C-EmGFP-GW/TOPO® Mam. Expr. Vector||Emerald GFP||C-terminal FP fusion||TOPO®||CMV|| ||bsd|
|Vivid Colors™ pcDNA™ 6.2/N-EmGFP-GW/TOPO® Mam. Expr. Vector||Emerald GFP||N-terminal FP fusion||TOPO®||CMV||N-term.V5||bsd|
|Vivid Colors™ pcDNA™ 6.2/C-YFP-GW/TOPO® Mam. Expr. Vector||Yellow||C-terminal FP fusion||TOPO®||CMV|| ||bsd|
|Vivid Colors™ pcDNA™ 6.2/N-YFP-GW/TOPO® Mam. Expr. Vector||Yellow||N-terminal FP fusion||TOPO®||CMV||N-term.V5||bsd|
|NT-GFP Fusion TOPO TA Expression Kit||Cycle 3 GFP||N-terminal FP fusion||TOPO®||CMV|| ||G418|
|CT-GFP Fusion TOPO TA Expression Kit||Cycle 3 GFP||C-terminal FP fusion||TOPO®||CMV|| ||G418|
|pcDNA™ -DEST47 Gateway® Vector||Cycle 3 GFP||C-terminal FP fusion||Gateway®||CMV|| ||G418|
|pcDNA™ -DEST53 Gateway® Vector||Cycle 3 GFP||N-terminal FP fusion||Gateway®||CMV|| ||G418|
|pBlue TOPO® TA Expression Kit||beta-gal||n/a||TOPO®||any||n/a||neo|
|pGlow TOPO® TA Expression Kit||Emerald GFP||n/a||TOPO®||any||n/a||neo|
|pTracer-CMV/Bsd||Cycle 3 GFP||native||R.E.||CMV|| ||GFP-bsd fusion|
|pTracer-CMV/Bsd Kit||Cycle 3 GFP||native||R.E.||CMV|| ||GFP-bsd fusion|
|pTracer-EF/Bsd A, B, & C||Cycle 3 GFP||native + tag||R.E.||EF-1 alpha||C-term.V5, 6xHis||GFP-bsd fusion|
|pTracer-EF/Bsd Kit||Cycle 3 GFP||native + tag||R.E.||EF-1 alpha||C-term.V5, 6xHis||GFP-bsd fusion|
|pTracer-CMV2 Kit||Cycle 3 GFP||native||R.E.||CMV|| ||GFP-zeo fusion|
|pTracer-CMV2 Vector||Cycle 3 GFP||native||R.E.||CMV|| ||GFP-zeo fusion|
|pTracer-SV40 Kit||Cycle 3 GFP||native||R.E.||SV40|| ||GFP-zeo fusion|
|pTracer-SV40 Vector||Cycle 3 GFP||native||R.E.||SV40|| ||GFP-zeo fusion|
|pTracer-EF Kit||Cycle 3 GFP||native + tag||R.E.||EF-1 alpha||C-term.V5, 6xHis||GFP-zeo fusion|
|pTracer-EF A, B, & C Vector||Cycle 3 GFP||native + tag||R.E.||EF-1 alpha||C-term.V5, 6xHis||GFP-zeo fusion|
|pRSET/CFP||Cyan||(n/a: bacterial expression clone)||6xHis, Xpress™, EK||n/a|
|pRSET/BFP||Blue||(n/a: bacterial expression clone)||6xHis, Xpress™, EK||n/a|
|pLenti6.2-GW/EmGFP Expression Control Vector||Emerald GFP||(GFP control only; no cloning site)||CMV||n/a||bsd|| |