pHcRed1 Vector
pHcRed1 is a bacterial expression vector which encodes the far-red fluorescent protein, HcRed1. HcRed1 was generated by mutagenesis of a non-fluorescent chromoprotein from the reef coral Heteractis crispa. The HcRed1 coding sequence has been human codon-optimized for higher expression in mammalian cells. The sequence upstream of HcRed1 has been converted to a Kozak consensus sequence to further increase the translation efficiency in eukaryotic cells.
The HcRed1 coding sequence is flanked by distinct multiple cloning sites (MCS) so that the gene can be easily excised from pHcRed1. Alternatively, the HcRed1 coding sequence can be amplified by PCR. In E. coli, HcRed1 is expressed from the lac promoter as a fusion with several additional amino acids, including the first five amino acids of the LacZ protein. Note, however, that if you excise the HcRed1 coding sequence using a restriction site in the 5' MCS, the resulting fragment will encode the native (i.e., non-fusion) HcRed1 protein. pHcRed1 is a pUC 19 derivative (pPD16.43) which provides a high-copy-number origin of replication as well as an ampicillin resistance gene for propagation and selection in E. coli.
Overview
- Far-red fluorescent protein, ideal for in vivo imaging
- HcRed1 fluorescent protein excitation and emission maxima: 588 and 618 nm, respectively
- HcRed1 is detected on a Western blot using anti-RCFP polyclonal pan antibody
Applications
- Fusions
- Protein localization studies
- General reporter for mammalian cells
- Monitoring transfection efficiencies
- In vivo imaging
Basic fluorescent protein vector map
Basic fluorescent protein vector map. Use this bacterial expression vector as a source of the fluorescent protein gene. Note: There is a stop codon at the 5' end of the 3' MCS. The 3' MCS should not be used for cloning.
Four-color visualization of Living Colors Fluorescent Proteins
Four-color visualization of Living Colors Fluorescent Proteins. HEK 293 clonal cell lines stably expressing AmCyan1, ZsGreen1, ZsYellow1, or HcRed1 were mixed and plated in the same culture dish and imaged at 20X magnification.
Excitation and emission spectra of reef coral fluorescent proteins AmCyan1, ZsGreen1, ZsYellow1, DsRed2, AsRed2, and HcRed1
Excitation and emission spectra of reef coral fluorescent proteins AmCyan1, ZsGreen1, ZsYellow1, DsRed2, AsRed2, and HcRed1.
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Basic fluorescent protein vector map
Basic fluorescent protein vector map. Use this bacterial expression vector as a source of the fluorescent protein gene. Note: There is a stop codon at the 5' end of the 3' MCS. The 3' MCS should not be used for cloning.
Four-color visualization of Living Colors Fluorescent Proteins
Four-color visualization of Living Colors Fluorescent Proteins. HEK 293 clonal cell lines stably expressing AmCyan1, ZsGreen1, ZsYellow1, or HcRed1 were mixed and plated in the same culture dish and imaged at 20X magnification.
Excitation and emission spectra of reef coral fluorescent proteins AmCyan1, ZsGreen1, ZsYellow1, DsRed2, AsRed2, and HcRed1
Excitation and emission spectra of reef coral fluorescent proteins AmCyan1, ZsGreen1, ZsYellow1, DsRed2, AsRed2, and HcRed1.
