Synthetic Gene DataBase
 

Synthetic Gene 66


 
  Welcome, Guest!

Field NameNatural GeneSynthetic Gene
SGDB Gene ID5966
GenBank AccessionL20587AY936883
GenBank GI46305761197044
Gene Namerevrev
Gene Length (bp)2639357
Specieshuman immunodeficiency virus (HIV1)Homo sapiens
StrainsType 1EBV-transformed lymphoblastoid cell line (BLCL) B157
CDSatggcaggaagaagcgaggacgaccagctgctgcaagccatccagatcataaagatcctg
taccaaagcagtaagtaacgctaatgcatcatagggacctgctagcaataataattatta
gtgctttgctgtttataaatgtaattttatgggggtttattcttagaaaatatttagaac
aaaaggaacaagacagaaaggagagagaaatacttgagaggttaagaagaataagagaaa
ttagggatgatagtgactatgaaagcaatggagaagaggaacaagaagttatggaccttg
tacttagccatggctttgataaccccatgtttgagccttagacagctatatgcaacagtc
tatgctggggtgcctgtatgggaagatgcaacaccagtactattctgtgcttcagatgct
aacctaacaagcactgaaaagcataatatttgggcatcacaagcctgtgttcctacagac
cccactccatatgaatatccattgcacaatgtgacagatgactttaatatatggaaaaat
tacatggtagaacaaatgcaggaagacattattagtttatgggaccagagtcttaaacct
tgtgttcaaatgactttcctgtgtgtacaaatggagtgtacaaacatagctggaacaaca
aatgaaaaccttatgaagaagtgtgagtttaatgtaaccactgttatcaaagacaaaaag
gagaaaaaacaggctctattctatgtatcagatttgatggaactgaatgagacaagcagc
acaaataagacaaacagcaaaatgtatacattaactaattgtaactccacaaccatcacg
caagcctgtccaaaggtatcttttgaaccaattccaatacactattgtgctccagcagga
tatgctatctttaagtgtaacagcacagaatttaatggaacaggcacatgcagaaacata
acggtagttacttgtacacatggcattaggccaacagtaagtactcagctaatattaaat
gggacactctctaaaggaaaaataagaatgatggcaaaagatattttggaaggtggaaaa
aatatcatagtgaccctaaactctaccctaaacatgacctgtgaaagaccacaaatagac
atacaagagatgagaataggtccaatggcctggtacagcatgggaatagggggaacagca
ggaaacagctcaagggcagcttattgcaagtataatgccactgattggggaaaaatatta
aaacaaacagctgaaaggtatttagaactagtaaacaatacaggtagtattaacatgaca
ttcaatcacagcagcggtggagatctagaggtaacccatttacactttaactgtcatgga
gaattcttttattgtaacacagctaagatgtttaattataccttttcatgtaacggaacc
acctgtagtgttagtaatgttagtcaaggtaacaatggcactctaccttgcaaactgaga
caggtggtaaggtcatggataaggggacagtcgggactctatgcacctcccatcaaaggt
aatctaacatgtatgtcaaacataactggaatgatcctacaaatggataacacatggaac
agcagcaacaacaatgtaacatttagaccaatagggggagacatgaaagatatatggaga
actgaattgttcaactacaaagtagtaagggtaaaaccttttagtgtggcacccacacgt
attgcaaggccagtcataagcactagaactcatagagaaaaaagagcagtaggattggga
atgctattcttgggggttctaagtgcagcaggtagcactatgggcgcagcggcaacaacg
ctggcggtacagacccacactttgctgaagggtatagtgcaacagcaggacaacctgcta
agagcaatacaggcccagcagcaattgctgaggctatctttatggggtatcagacaactc
cgagctcgcctgctagccttagaaaccttactacagaatcagcaactcctaagcctatgg
ggctgtaaaggaaagctagtctgctacacatcagtaaaatggaatagaacatggatagga
aacgaaagcatttgggacaccttaacatggcaggaatgggatcggcagataagcaacata
agctccaccatatatgaggaaatacaaaaggcacaagtacagcaggaacaaaatgagaaa
aagttgctggagttagatgaatgggcctctatttggaattggcttgacataactaaatgg
ttgtggtatataaaaatagcaataatcatagtaggagcactagtaggggtgagagttatc
atgatagtacttaatatagtgaaaaacattaggcagggatatcaacccctctcgttacag
atccccaaccatcaccaagaggaagcaggaacgccaggaagaacaggaggaggaggtgga
gaagaaggcaggcccaggtggataccctcgccgcaagggttcttgccactgttgtacacg
gacctcagaacaataatattgtggacttaccacctcttgagcaacttagcatcagggatc
cagaaggtgatcagctatctgaggcttggactgtggatcctagggcagaagataattaa
atggccggcagaagcggcgacagcgacgaggagctgctgaaaaccgtgcggctcatcaag
ttcctgtaccagagcaaccctccacccagccccgagggcaccagacaggcccggagaaac
cggaggaggcggtggagagagaggcagcggcagatcagaagcatcagcgagcggattctg
agcacctacctgggcagaagcgccgagcccgtgcccctgcagctgccccccctggagaga
ctgaccctggactgcaatgaggattgcggcaccagcggcacccagggcgtgggcagcccc
cagatcctggtggagagccctgccgtgctggagagcggcaccaaggaagatctgtga
5' End
3' End
NotesNo information on the natural rev gene in the paper so the rev gene in HIV1 genome (L20587) was given for reference.regulator of virus expression
Expression VectorNApEGFP-N1
Assay MethodsNAFluorescence dection
ResultsNot reported50-60% of cells are GFP-positive
Protein Function
Recoding PurposeTo improve expression
Synthesized ByGeneArt
Recoding Methodcodon optimized for Homo sapiens
Publication Author(s)van Baalen CA, Kwa D, Verschuren EJ, Reedijk ML, Boon AC, de Mutsert G, Rimmelzwaan GF, Osterhaus AD, Gruters RA.
Corresponding AuthorAlbert D.M.E. Osterhaus
Corresponding AddressDepartment of Virology, Erasmus MC, University Medical Center and Postgraduate School of Molecular Medicine, Rotterdam, The Netherlands.
Publication Year2005
Publication TitleFluorescent antigen-transfected target cell cytotoxic T lymphocyte assay for ex vivo detection of antigen-specific cell-mediated cytotoxicity.
AbstractEx vivo detection of virus-specific cytotoxic T lymphocyte (CTL) responses is limited to the use of methods assessing cytokine production, degranulation, or perforin contents of antigen-specific CD8+ T cells. Generally, their cytotoxic activity is detectable only after cultivation. We describe the fluorescent antigentransfected target cellCTL (FATT-CTL) assay, which measures antigen-specific cytotoxicity ex vivo. Target cells were generated by nucleofection with DNA vectors encoding antigengreen fluorescent protein (GFP) fusion proteins. After coculture at various effector : target (E : T) cell ratios, viable and dead GFP-positive cells were quantified by flow cytometry, and antigen-specific target-cell elimination was calculated. The assay was validated with human immunodeficiency virus (HIV) and influenza virusspecific CTL clones and revealed cytotoxicity at lower E : T cell ratios than standard 51Cr-release assays. Moreover, antigen-specific cytotoxicity was detected ex vivo within 1 day in peripheral blood mononuclear cells from HIV-infected individuals. The FATT-CTL assay provides a versatile tool that will advance our understanding of cell-mediated immunity.
JournalJ Infect Dis.. 192(7): 1183-90.
SummaryFour HIV1 genes were codon optimized for human (by GeneArt) and conjugated with GFP to act as markers in ex vivo dection of virus-specific cytotoxic T lymphocyte (CTL) response. They appeared to work well as expected.
Comments
Discussion http://www.evolvingcode.net/forum/viewtopic.php?t=513
PubMed ID16136460
Submitter NameWu, Gang
Submitter AddressDepartment of Biological Sciences, University of Maryland Baltimore County, 1000 Hilltop Circle, Baltimore, MD 21250 USA
Entry ConfirmationNo
 
 

Copyright 2004 the Freeland Bioinformatics Lab, All Rights Reserved. | Contact Us | About this site