pDsRed-Monomer-Hyg-N1 Vector

Description

pDsRed-Monomer-Hyg-N1 encodes DsRed-Monomer, a monomeric mutant of theDiscosomasp. red fluorescent protein DsRed. pDsRed-Monomer-Hyg-N1 is designed for studies in mammalian systems. This vector allows expression of a protein of interest as an N-terminal fusion to DsRed-Monomer. The vector contains a hygromycin resistance gene for the selection of stable transfectants. pDsRed-Monomer-Hyg-N1 can also be used as a cotransfection marker since the unmodified vector constitutively expresses DsRed-Monomer. The fluorescent protein coding sequence in this construct has been human codon-optimized for efficient expression and enhanced brightness in mammalian cells. Our products are to be used forResearch Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Back Schematic for fluorescent fusion protein vectors.Panel A.N-terminal fluorescent protein FP vector.Panel B.C-terminal fluorescent protein vector. Back 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. Back Back DsRed-Monomer is a monomeric protein.Panel A. Recombinant DsRed-Express and DsRed-Monomer fluorescent proteins 100 mug were analyzed by FPLC gel filtration chromatography. Overall absorbance A280 and chromophore excitation A557 of the eluted material were monitored simultaneously. DsRed-Monomer elutes from the column at a retention time 39 min corresponding to a molecular weight of 28 kDa. The calculated molecular weight of DsRed-Monomer is 26.8 kDa. DsRed-Express is a tetrameric protein that elutes at an earlier retention time 33 min corresponding to a molecular weight of 89 kDa.Panel B. Pseudonative gel analysis of proteins. The oligomeric structure of proteins is preserved during SDS PAGE analysis if samples are kept at 4C and not boiled prior to loading on a gel. Boiled and unboiled recombinant proteins 7.5 mug were separated by SDS PAGE electrophoresis 12 % acrylamide . In both the boiled denatured and unboiled nondenatured samples, DsRed-Monomer fluorescent protein runs as a uniform band of 30 kDa due to its monomeric structure. The unboiled nondenatured DsRed-Express runs at a much higher molecular weight than its boiled denatured counterpart due to its tetrameric structure. Back