Synthetic Gene DataBase
 

Synthetic Gene 44


 
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Field NameNatural GeneSynthetic Gene
SGDB Gene ID3844
GenBank AccessionAF168419AF506027
GenBank GI710573321464837
Gene NameDsRed (drFP583)mRFP1
Gene Length (bp)678678
SpeciesDiscosoma sp (coral)Homo sapiens
StrainsHela Cells
CDSatgaggtcttccaagaatgttatcaaggagttcatgaggtttaaggttcgcatggaagga
acggtcaatgggcacgagtttgaaatagaaggcgaaggagaggggaggccatacgaaggc
cacaataccgtaaagcttaaggtaaccaaggggggacctttgccatttgcttgggatatt
ttgtcaccacaatttcagtatggaagcaaggtatatgtcaagcaccctgccgacatacca
gactataaaaagctgtcatttcctgaaggatttaaatgggaaagggtcatgaactttgaa
gacggtggcgtcgttactgtaacccaggattccagtttgcaggatggctgtttcatctac
aaggtcaagttcattggcgtgaactttccttccgatggacctgttatgcaaaagaagaca
atgggctgggaagccagcactgagcgtttgtatcctcgtgatggcgtgttgaaaggagag
attcataaggctctgaagctgaaagacggtggtcattacctagttgaattcaaaagtatt
tacatggcaaagaagcctgtgcagctaccagggtactactatgttgactccaaactggat
ataacaagccacaacgaagactatacaatcgttgagcagtatgaaagaaccgagggacgc
caccatctgttcctttaa
atggcctcctccgaggacgtcatcaaggagttcatgcgcttcaaggtgcgcatggagggc
tccgtgaacggccacgagttcgagatcgagggcgagggcgagggccgcccctacgagggc
acccagaccgccaagctgaaggtgaccaagggcggccccctgcccttcgcctgggacatc
ctgtcccctcagttccagtacggctccaaggcctacgtgaagcaccccgccgacatcccc
gactacttgaagctgtccttccccgagggcttcaagtgggagcgcgtgatgaacttcgag
gacggcggcgtggtgaccgtgacccaggactcctccctgcaggacggcgagttcatctac
aaggtgaagctgcgcggcaccaacttcccctccgacggccccgtaatgcagaagaagacc
atgggctgggaggcctccaccgagcggatgtaccccgaggacggcgccctgaagggcgag
atcaagatgaggctgaagctgaaggacggcggccactacgacgccgaggtcaagaccacc
tacatggccaagaagcccgtgcagctgcccggcgcctacaagaccgacatcaagctggac
atcacctcccacaacgaggactacaccatcgtggaacagtacgagcgcgccgagggccgc
cactccaccggcgcctaa
5' End
3' End
Notes
Expression VectorNApcDNA3
Assay MethodsNAFluorescence imaging
ResultsExpression not determinedWithin 12h cells displayed strong red fluorescence evenly distributed thoughout the nucleus and cytoplasm.
Protein FunctionReporter gene (Red fluorescent protein FP583)
Recoding PurposeTo improve expression
Synthesized ByAuthors
Recoding MethodCodon usage optimized for human codon preferences
Publication Author(s)Campbell RE, Tour O, Palmer AE, Steinbach PA, Baird GS, Zacharias DA, Tsien RY.
Corresponding AuthorRoger Y. Tsien
Corresponding AddressDepartment of Pharmacology. University of California at San Diego, 9500 Gilman Drive, La Jolla, CA 92093, USA.
Publication Year2002
Publication TitleA monomeric red fluorescent protein.
AbstractAll coelenterate fluorescent proteins cloned to date display some form of quaternary structure, including the weak tendency of Aequorea green fluorescent protein (GFP) to dimerize, the obligate dimerization of Renilla GFP, and the obligate tetramerization of the red fluorescent protein from Discosoma (DsRed). Although the weak dimerization of Aequorea GFP has not impeded its acceptance as an indispensable tool of cell biology, the obligate tetramerization of DsRed has greatly hindered its use as a genetically encoded fusion tag. We present here the stepwise evolution of DsRed to a dimer and then either to a genetic fusion of two copies of the protein, i.e., a tandem dimer, or to a true monomer designated mRFP1 (monomeric red fluorescent protein). Each subunit interface was disrupted by insertion of arginines, which initially crippled the resulting protein, but red fluorescence could be rescued by random and directed mutagenesis totaling 17 substitutions in the dimer and 33 in mRFP1. Fusions of the gap junction protein connexin43 to mRFP1 formed fully functional junctions, whereas analogous fusions to the tetramer and dimer failed. Although mRFP1 has somewhat lower extinction coefficient, quantum yield, and photostability than DsRed, mRFP1 matures >10 times faster, so that it shows similar brightness in living cells. In addition, the excitation and emission peaks of mRFP1, 584 and 607 nm, are approximately 25 nm red-shifted from DsRed, which should confer greater tissue penetration and spectral separation from autofluorescence and other fluorescent proteins.
JournalProc Natl Acad Sci U S A.. 99(12): 7877-82.
SummaryThree versions of DsRed gene were designed with reference to the codon usage in human and expressed in human HeLa cells. All recoded genes showed strong red fluorescence in 12h.
CommentsThe study itself focused more on the activity of proteins coded by recoded genes.
Discussion http://www.evolvingcode.net/forum/viewtopic.php?t=500
PubMed ID12060735
Submitter NameWu, Gang
Submitter AddressDepartment of Biological Sciences, University of Maryland Baltimore County, 1000 Hilltop Circle, Baltimore, MD 21250 USA
Entry ConfirmationNo
 
 

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