Synthetic Gene DataBase

Synthetic Gene 68

  Welcome, Guest!

Field NameNatural GeneSynthetic Gene
SGDB Gene ID6068
GenBank AccessionK03455AY936885
GenBank GI190638261197048
Gene Nametattat
Gene Length (bp)261309
Specieshuman immunodeficiency virus (HIV1)Homo sapiens
StrainsType 1EBV-transformed lymphoblastoid cell line (BLCL) B157
5' End
3' End
NotesNo wild-type gene information mentioned in the paper. So the HXB2 strain tat gene was collected. Need to contact the authors for confirmationmutated for transactivation nuclear localization and integrin receptor binding; codon optimized for Homo sapiens
Expression VectorNApEGFP-N1
Assay MethodsNAFluorescence dection
ResultsNot reported50-60% of cells are GFP-positive
Protein Functiontransactivator protein
Recoding PurposeTo improve expression
Synthesized ByGeneArt
Recoding Methodcodon optimized for Homo sapiens. Done by GeneArt. Also mutated for transactivation nuclear
localization and integrin receptor binding.
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.
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