DsRed-Express and DsRed-Express2 fluorescent proteins

pDsRed-Express-N1 encodes DsRed-Express, a variant of Discosoma sp. red fluorescent protein (DsRed). DsRed-Express contains nine amino acid substitutions (as compared to DsRed1), which improve the solubility, increase the maturation rate, and decrease the level of residual green emission. pDsRed-Express allows genes cloned into the multiple cloning site (MCS) upstream of the DsRed-Express coding sequence to be expressed as fusions to the N-terminus of DsRed-Express. The unmodified vector will express DsRed-Express in mammalian cells.

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Schematic for fluorescent fusion protein vectors

Schematic for fluorescent fusion protein vectors. Panel A. N-terminal fluorescent protein (FP) vector. Panel B. C-terminal fluorescent protein vector.

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Hygromycin DsRed-Monomer and AcGFP1 expression vectors

Hygromycin DsRed-Monomer and AcGFP1 expression vectors. Vector maps of our DsRed-Monomer and pAcGFP1 Hyg-N1 (Panel A) and Hyg-C1 (Panel B) mammalian expression vectors for fusion applications. Each vector contains a hygromycin selection cassette. These vectors can be used together with our standard N1/C1 vectors containing the neomycin selection cassette to establish double stable cell lines expressing two different Living Colors proteins.

pDsRed-Express2-C1 encodes DsRed-Express2, a variant of Discosoma sp. red fluorescent protein. It can be used to construct fusions to the C-terminus of DsRed-Express2 which retain the fluorescent properties of the native DsRed-Express2 protein. Its expression can be monitored by flow cytometry, and its intracellular localization can be determined by fluorescence microscopy. pDsRed-Express2-C1 can also be used as a cotransfection marker; the unmodified vector will express DsRed-Express2.

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Schematic for fluorescent fusion protein vectors

Schematic for fluorescent fusion protein vectors. Panel A. N-terminal fluorescent protein (FP) vector. Panel B. C-terminal fluorescent protein vector.

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Robust expression of DsRed-Express2 in mouse bone marrow hematopoietic stem and progenitor cells

Robust expression of DsRed-Express and DsRed-Express2 in mouse bone marrow hematopoietic stem and progenitor cells. Mononuclear bone marrow cells were transduced with retroviral vectors encoding DsRed-Express (Panel A), DsRed-Express2 (Panel B), or EGFP (Panel C), and fluorescent cells were sorted 87 hr later. Red and green fluorescence signals were detected using the PE and FITC filter sets, respectively. The lines represent gates defined by analyzing untransduced cells.

pDsRed-Express-1 is a promoterless mammalian expression vector that can be used to monitor transcription from different promoters and promoter/enhancer combinations inserted into MCS. It encodes DsRed-Express, a variant of Discosoma sp. red fluorescent protein (DsRed). DsRed-Express contains nine amino acid substitutions which improve the solubility of the protein, reduce the time from transfection to detection of red fluorescence, and decrease the level of residual green emission. When DsRed-Express is expressed in mammalian cell cultures, red-emitting cells can be detected by either fluorescence microscopy or flow cytometry 8–12 hours after transfection. Although DsRed-Express most likely forms the same tetrameric structure as wild-type DsRed, DsRed-Express displays a reduced tendency to aggregate. The DsRed-Express coding sequence is human codon-optimized for high expression in mammalian cells.

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Vector map for promoterless fluorescent protein vectors AcGFP1-1, DsRed2-1, DsRed-Express-1, HcRed1-1 and ZsGreen1-1

Vector map for promoterless fluorescent protein vectors pAcGFP1-1, pDsRed2-1, pDsRed-Express-1, pHcRed1-1 and pZsGreen1-1.

pCMV-DsRed-Express is a mammalian expression vector designed for use as a cotransfection marker, together with an expression construct of interest. It constitutively expresses the red fluorescent protein DsRed-Express, which can be detected by fluorescence microscopy to provide direct visual evidence of transfection. Cells can also be sorted by flow cytometry to enrich for transfected cells.

pCMV-DsRed-Express Vector can be transfected into mammalian cells using any standard transfection method. If required, stable transfectants can be selected using G418.

This lentiviral expression vector encodes a DsRed-Express2 fluorescent protein tag. The protein is a rapidly maturing tetrameric fluorescent protein from Discosoma sp. that has minimal cytotoxicity, high solubility, and exhibits strong and stable expression mammalian cells. Inserting a cDNA in the MCS upstream of the DsRed-Express2 coding sequence joins your protein of interest to the N-terminus of the tag, and allows the fusion protein to be tracked and studied in transduced cells. To package the vector into high-titer, replication-incompetent lentivirus, we recommend using the Lenti-X Packaging Single Shots and the Lenti-X 293T Cell Line. The resulting lentivirus can then be used to transduce virtually any mammalian cell type.

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Lentiviral vectors with fluorescent proteins

Lentiviral vectors with fluorescent proteins. Lenti-X vectors contain sequence elements that facilitate lentiviral packaging and/or boost transgene expression, including the LTRs, packaging signal (Ψ), Rev response element (RRE), and central polypurine tract/central termination sequence (cPPT/CTS) from HIV-1; and the woodchuck hepatitis virus post-transcriptional regulatory element (WPRE). Vectors can express your protein fused at its N- or C- terminus to either a green (AcGFP1) or red (DsRed-Monomer) fluorescent protein tag, or coexpress it as a separate protein along with ZsGreen1 (shown), mCherry, or tdTomato.

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Robust expression of DsRed-Express2 in mouse bone marrow hematopoietic stem and progenitor cells

Robust expression of DsRed-Express and DsRed-Express2 in mouse bone marrow hematopoietic stem and progenitor cells. Mononuclear bone marrow cells were transduced with retroviral vectors encoding DsRed-Express (Panel A), DsRed-Express2 (Panel B), or EGFP (Panel C), and fluorescent cells were sorted 87 hr later. Red and green fluorescence signals were detected using the PE and FITC filter sets, respectively. The lines represent gates defined by analyzing untransduced cells.