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dc.contributor.authorVider, Jelenaen_US
dc.date.accessioned2012-04-11en_US
dc.date.accessioned2012-07-12T23:15:33Z
dc.date.accessioned2017-03-01T22:51:50Z
dc.date.available2017-03-01T22:51:50Z
dc.date.issued2003en_US
dc.date.modified2012-07-12T23:15:33Z
dc.identifier.issn1522-8002en_US
dc.identifier.urihttp://hdl.handle.net/10072/45828
dc.description.abstractTo optimize the sensitivity of imaging HSV1-tk/GFP reporter gene expression, a series of HSV1-tk/GFP mutants was developed with altered nuclear localization and better cellular enzymatic activity, compared to that of the native HSV1-tk/GFP fusion protein (HSV1-tk/GFP). Several modifications of HSV1-tk/GFP reporter gene were performed, including targeted inactivating mutations in the nuclear localization signal (NLS), the addition of a nuclear export signal (NES), a combination of both mutation types, and a truncation of the first 135 bp of the native hsv1-tk coding sequence containing a "cryptic" testicular promoter and the NLS. A recombinant HSV1-tk/GFP protein and a highly sensitive sandwich enzyme-linked immunosorbent assay for HSV1-tk/GFP were developed to quantitate the amount of reporter gene product in different assays to allow normalization of the data. These different mutations resulted in various degrees of nuclear clearance, predominant cytoplasmic distribution, and increased total cellular enzymatic activity of the HSV1-tk/GFP mutants, compared to native HSV1-tk/GFP when expressed at the same levels. This appears to be the result of improved metabolic bioavailability of cytoplasmically retargeted mutant HSV1-tk/GFP enzymes for reaction with the radiolabeled probe (e.g., FIAU). The analysis of enzymatic properties of different HSV1-tk/GFP mutants using FIAU as a substrate revealed no significant differences from that of the native HSV1-tk/GFP. Improved total cellular enzymatic activity of cytoplasmically retargeted HSV1-tk/GFP mutants observed in vitro was confirmed by noninvasive imaging of transduced subcutaneous tumor xenografts bearing these reporters using [(131)I]FIAU and a gamma-camera.en_US
dc.description.peerreviewedYesen_US
dc.description.publicationstatusYesen_US
dc.format.extent1436152 bytes
dc.format.mimetypeapplication/pdf
dc.publisherNeoplasia Pressen_US
dc.publisher.placeUnited States of Americaen_US
dc.publisher.urihttp://www.neoplasia.com/abstract.php?msid=190en_US
dc.relation.ispartofpagefrom245en_US
dc.relation.ispartofpageto254en_US
dc.relation.ispartofissue3en_US
dc.relation.ispartofjournalNeoplasiaen_US
dc.relation.ispartofvolume5en_US
dc.subject.fieldofresearchMolecular Targetsen_US
dc.subject.fieldofresearchcode111207en_US
dc.titleCytoplasmically retargeted HSV1-tk/GFP reporter gene mutants for optimization of noninvasive molecular-genetic imaging.en_US
dc.typeJournal article
dc.type.descriptionJournal Articles (Refereed Article)en_US
dc.type.codec1xen_US
gro.facultyGriffith Health Facultyen_US
gro.rights.copyrightCopyright 2003 Neoplasia Press. The attached file is reproduced here in accordance with the copyright policy of the publisher. Please refer to the journal's website for access to the definitive, published version.en_US
gro.date.issued2003
gro.hasfulltextFull Text


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