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<div class="pre-content"><div><div class="bk_prnt"><p class="small">NCBI Bookshelf. A service of the National Library of Medicine, National Institutes of Health.</p><p>Molecular Imaging and Contrast Agent Database (MICAD) [Internet]. Bethesda (MD): National Center for Biotechnology Information (US); 2004-2013. </p></div><div class="iconblock clearfix whole_rhythm no_top_margin bk_noprnt"><a class="img_link icnblk_img" title="Table of Contents Page" href="/books/n/micad/"><img class="source-thumb" src="/corehtml/pmc/pmcgifs/bookshelf/thumbs/th-micad-lrg.png" alt="Cover of Molecular Imaging and Contrast Agent Database (MICAD)" height="100px" width="80px" /></a><div class="icnblk_cntnt eight_col"><h2>Molecular Imaging and Contrast Agent Database (MICAD) [Internet].</h2><a data-jig="ncbitoggler" href="#__NBK23131_dtls__">Show details</a><div style="display:none" class="ui-widget" id="__NBK23131_dtls__"><div>Bethesda (MD): <a href="https://www.ncbi.nlm.nih.gov/" ref="pagearea=page-banner&targetsite=external&targetcat=link&targettype=publisher">National Center for Biotechnology Information (US)</a>; 2004-2013.</div></div><div class="half_rhythm"><ul class="inline_list"><li style="margin-right:1em"><a class="bk_cntns" href="/books/n/micad/">Contents</a></li></ul></div><div class="bk_noprnt"><form method="get" action="/books/n/micad/" id="bk_srch"><div class="bk_search"><label for="bk_term" class="offscreen_noflow">Search term</label><input type="text" title="Search this book" id="bk_term" name="term" value="" data-jig="ncbiclearbutton" /> <input type="submit" class="jig-ncbibutton" value="Search this book" submit="false" style="padding: 0.1em 0.4em;" /></div></form></div></div><div class="icnblk_cntnt two_col"><div class="pagination bk_noprnt"><a class="active page_link prev" href="/books/n/micad/sosgreenfluorescencepro2/" title="Previous page in this title">< Prev</a><a class="active page_link next" href="/books/n/micad/soshilytefluor647/" title="Next page in this title">Next ></a></div></div></div></div></div>
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<div class="main-content lit-style" itemscope="itemscope" itemtype="http://schema.org/CreativeWork"><div class="meta-content fm-sec"><h1 id="_NBK23131_"><span class="title" itemprop="name">HSV1-TK/GFP/Fluc</span></h1><div itemprop="alternativeHeadline" class="subtitle whole_rhythm">TGL</div><p class="contrib-group"><span itemprop="author">Huiming Zhang</span>, PhD.</p><a data-jig="ncbitoggler" href="#__NBK23131_ai__" style="border:0;text-decoration:none">Author Information and Affiliations</a><div style="display:none" class="ui-widget" id="__NBK23131_ai__"><div class="contrib half_rhythm"><span itemprop="author">Huiming Zhang</span>, PhD<div class="affiliation small">
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National Center for Biotechnology Information, NLM, NIH, Bethesda, MD,
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||
<span class="before-email-separator"></span><span class="email-label">Email: </span><a href="mailto:dev@null" data-email="vog.hin.mln.ibcn@dacim" class="oemail">vog.hin.mln.ibcn@dacim</a>
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||
</div></div></div><p class="small">Created: <span itemprop="datePublished">October 21, 2008</span>; Last Update: <span itemprop="dateModified">December 1, 2008</span>.</p></div><div class="jig-ncbiinpagenav body-content whole_rhythm" data-jigconfig="allHeadingLevels: ['h2'],smoothScroll: false" itemprop="text"><div id="TGL.T1" class="table"><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK23131/table/TGL.T1/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__TGL.T1_lrgtbl__"><table><tbody><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
|
||
<b>Chemical name:</b>
|
||
</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">HSV1-TK/GFP/Fluc</td><td rowspan="9" colspan="1" style="text-align:left;vertical-align:middle;"></td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
|
||
<b>Abbreviated name:</b>
|
||
</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">TGL</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
|
||
<b>Synonym:</b>
|
||
</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">TGL triple reporter</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
|
||
<b>Agent category:</b>
|
||
</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Protein</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
|
||
<b>Target:</b>
|
||
</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Other</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
|
||
<b>Target category:</b>
|
||
</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Other – gene expression</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
|
||
<b>Method of detection:</b>
|
||
</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Optical imaging, gamma imaging, SPECT</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
|
||
<b>Source of signal/contrast:</b>
|
||
</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Green fluorescence protein (GFP), firefly luciferase, and <sup>131</sup>I-FIAU</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
|
||
<b>Activation:</b>
|
||
</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Yes</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
|
||
<b>Studies:</b>
|
||
</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
|
||
<ul class="simple-list"><li class="half_rhythm"><div>
|
||
<img alt="Checkbox" src="/corehtml/pmc/css/bookshelf/2.26/img/studies.checkbox.png" />
|
||
<i>In vitro</i>
|
||
|
||
</div></li></ul>
|
||
<ul class="simple-list"><li class="half_rhythm"><div>
|
||
<img alt="Checkbox" src="/corehtml/pmc/css/bookshelf/2.26/img/studies.checkbox.png" /> Rodents
|
||
</div></li></ul>
|
||
</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">No structure is currently available in <a href="http://pubchem.ncbi.nlm.nih.gov" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">PubChem</a>.</td></tr></tbody></table></div></div><div id="TGL.Background"><h2 id="_TGL_Background_">Background</h2><p>[<a href="/sites/entrez?Db=pubmed&Cmd=DetailsSearch&Term=HSV1-tk%20+%20GFP%20+%20luciferase" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">PubMed</a>]</p><p>Reporter genes, also known as marker genes, possess a measurable phenotype distinguishable from the background of endogenous proteins (<a class="bk_pop" href="#TGL.REF.1">1</a>). Several reporter genes express proteins that can generate signals for <i>in vivo</i> imaging, such as the herpes simplex virus type 1 thymidine kinase (HSV1-TK) gene, the green fluorescence protein (GFP) gene, and the firefly luciferase (Fluc) gene (<a class="bk_pop" href="#TGL.REF.2">2</a>). The reporter gene is constructed with a “constitutive” promoter for continuous transcriptions or with an “inducible” promoter for controlled transcriptions (<a class="bk_pop" href="#TGL.REF.3">3</a>). Both types of gene constructs have been used in the expression of exogenous genes and/or endogenous genes to monitor the levels of gene delivery and the efficiency in cell/tissue transduction in gene therapy.</p><p>HSV1-TK/GFP/Fluc (TGL, Mw = 130 kDa) is a triple reporter protein used for multimodal <i>in vivo</i> imaging of gene expression (<a class="bk_pop" href="#TGL.REF.4">4</a>) with single photon emission computed tomography (SPECT) or fluorescence and bioluminescence imaging. TGL is produced from the expression of a triple-fusion reporter gene (TGL gene) in which transcription and translation are processed through a single open reading frame. TGL consists of three fused protein subunits: HSV1-TK, GFP, and Fluc, where polylysine is used as a linker to connect the protein subunits and to maintain the molecular stability in the fused protein. Each subunit corresponds to a specific imaging modality. HSV1-TK is a homodimer composed of two 376-residue subunits (<a class="bk_pop" href="#TGL.REF.5">5</a>), and it functions as a key enzyme in the pyrimidine-salvage pathway to catalyze the phosphorylation of thymidine (dT) to thymidine monophosphate (dTMP) in the presence of ATP and Mg<sup>2+</sup>. This feature has been used to design antiviral nucleoside analogs (i.e., ganciclovir) for antiviral therapy (<a class="bk_pop" href="#TGL.REF.5">5</a>) and radiolabeled substrates such as radiolabeled 2’-fluoro-2’-deoxy-1-β-D-arabino-furanosyl-5-iodo-uracil (FIAU) for SPECT imaging (<a class="bk_pop" href="#TGL.REF.6">6</a>). HSV can easily infect a variety of cells in that the expressed HSV1-TK can induce significant cytotoxicity in the presence of nucleoside analogs such as ganciclovir (<a class="bk_pop" href="#TGL.REF.7">7</a>). For this reason, HSV1-TK gene is also widely used as a suicide gene in cancer treatment. <sup>131</sup>I-Labeled FIAU is an active radiolabeled substrate of HSV1-TK that is detectable with SPECT (i.e., <sup>131</sup>I emits gamma-ray at 364 keV (81% abundance) with a half-life time of 8.02 days, which can be detected with a gamma camera or SPECT. GFP is a fluorescent protein of 238 amino acids that emits a bright green fluorescence (λ<sub>max</sub> = 509 nm) when illuminated with a blue light (λ<sub>max</sub> = 395 nm) (<a class="bk_pop" href="#TGL.REF.8">8</a>). The presence of the GFP subunit allows <i>in vitro</i>/<i>in vivo</i> fluorescence imaging and cell sorting with the fluorescence-activated cell sorting (FACS) technique. Fluc is an oxygenase (Mw = 62 kDa) extracted from <i>Photinus pyralis</i> (<a class="bk_pop" href="#TGL.REF.9">9</a>). In the presence of adenosine triphosphate (ATP) and O<sub>2</sub>, Fluc oxidizes the heterocyclic substrate <span class="small-caps">d</span>-luciferin to oxyluciferin and emits light in the wavelength range of 400–620 nm (<a class="bk_pop" href="#TGL.REF.10">10</a>). The Fluc subunit allows for planar whole-body bioluminescence imaging of gene-transduced cells.</p></div><div id="TGL.Synthesis"><h2 id="_TGL_Synthesis_">Synthesis</h2><p>[<a href="/sites/entrez?Db=pubmed&Cmd=DetailsSearch&Term=(%20HSV1-tk%20+%20GFP%20+%20luciferase)+AND+synthesis%0D%0A" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">PubMed</a>]</p><p>Ponomarev et al. described the preparation of the HSV1-TK/GFP/Fluc expression vector in multiple steps (<a class="bk_pop" href="#TGL.REF.4">4</a>). First, a construct SFG-HSV1-TK/GFP was obtained (<a class="bk_pop" href="#TGL.REF.11">11</a>). Plasmid pLTKRNL was digested with restriction enzyme <i>Bam</i>HI to generate a 2.2-kb fragment that encoded the open reading frame of HSV1-TK. This fragment was ligated into plasmid pcDNA.1/Zeo to result in the 7.8-kb plasmid pTKZeo3.2; this ligation was followed by sequential digestions with <i>Bam</i>HI and <i>Xma</i>l to produce a 1.2-kb TK fragment. At the same time, plasmid pEGFP-N1 was digested sequentially with <i>BgI</i>II and <i>Xma</i>l to generate a 4.7-kb DNA fragment containing the open reading frame of GFP. The 1.2-kb fragment was then ligated with the 4.7-kb fragment to form the 5.9-kb expression plasmid pTKGFP, in which the TK-GFP cDNA was under control of the cytomegalovirus immediately early (IE) promoter. After the ligation, the plasmid was cloned into a retroviral vector, the Moloney murine leukemia virus-based SFG vector, to yield SFG-TK-GFP. Second, two modified constructs, SFG-NES-HSV1-TK/GFP and SFG-Δ45-HSV1-TK/GFP, were obtained (<a class="bk_pop" href="#TGL.REF.6">6</a>). To generate SFG-NES-HSV1-TK/GFP, the Xenopus cDNA for encoding the 20 amino acids in the nuclear export signal (NES) sequence of the mitogen-activated protein (MAP) kinase kinase (MAPKK) was inserted into the N-terminus of HSV1-TK/GFP between His-9 and Ala-10 using <i>Mlu</i>L. To generate SFG-Δ45-HSV1-TK/GFP, the Δ45-HSV1 cDNA lacking the first 45 amino acids (135 nucleotides) of the original HSV1-TK was amplified with selective primers and subsequently cloned between the <i>Nco</i>l sites of SFG-HSV1-TK/GFP. Third, the commercially available cDNA of fused <i>Aequorea victoria</i> eGFP and Fluc (eGFP/Fluc) was obtained from the plasmid pEGF/Pluc and subcloned into the retroviral vector SFG-Ntp to yield the construct SFG-GL (<a class="bk_pop" href="#TGL.REF.4">4</a>). Finally, SFG-NES-HSV1-TK/GFP or SFG-Δ45-HSV1-TK/GFP was treated with <i>Nco</i>l and the fragment containing NES-HSV1-TK or Δ45-HSV1-TK was subcloned into the <i>Nco</i>I site of the SFG-GL to produce SFG-NES-HSV1-TK/GFP/Fluc (SFG-NES-TGL) or SFG-Δ45-HSV1-TK/GFP/Fluc (SFG-Δ45-TGL) (<a class="bk_pop" href="#TGL.REF.4">4</a>). [<sup>131</sup>I]-FIAU with >97% radiochemical purity was prepared by the iododestannylation reaction using the tin precursor and carrier-free <sup>131</sup>I.</p></div><div id="TGL.In_Vitro_Studies_Tes"><h2 id="_TGL_In_Vitro_Studies_Tes_"><i>In Vitro</i> Studies: Testing in Cells and Tissues</h2><p>[<a href="/sites/entrez?Db=pubmed&Cmd=DetailsSearch&Term=(%20HSV1-tk%20+%20GFP%20+%20luciferase)+AND+%22in+vitro%22" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">PubMed</a>]</p><p>Ponomarev et al. assessed the integrity of the fused reporter proteins with Western blot analysis (<a class="bk_pop" href="#TGL.REF.4">4</a>). The molecular weight of the NES-TGL and Δ45-TGL proteins as determined with Western blot analysis matched the predicted molecular weights. These proteins were further examined with anti-HSV1-TK, anti-GFP, and anti-Fluc antibodies to demonstrate the coexistence of the triple subunits in the full reporter proteins. Ponomarev et al. then imaged the cytoplasmic localizations of the NES-TGL and Δ45-TGL reporter proteins with fluorescence microscopy (<a class="bk_pop" href="#TGL.REF.4">4</a>). Human glioma U87 cells were transduced with the retroviral vectors SFG-NES-TGL or SFG-Δ45-TGL <i>in vitro</i>, followed by a bulk culture for 48 h. The obtained U87-NES-TGL cells or U87-Δ45-TGL cells were sorted for positive GFP expression using FACS with 488-nm excitation beam and 510-nm emission filter. The subcellular localization of NES-TGL and Δ45-TGL was visualized with fluorescence microscopy with similar excitation emission parameters. The GFP/Fluc signal appeared to be lower in the nucleus than in cytoplasm, but it was still detectable. Reporter gene expression in these transduced U87 cells was quantified with FACS, a bioluminescence assay, and a radiotracer [<sup>14</sup>C]-FIAU assay. The level of GFP fluorescence was found to be ~1.3 × 10<sup>3</sup> relative fluorescent units (RU) in the U87-NES-TGL cells and ~2.1 × 10<sup>3</sup> RU in the U87-Δ45-TGL cells. The influx rate constant <i>Ki</i> of <sup>14</sup>C-labeled FIAU was ~1.4 media ml/min per g in the U87-NES-TGL cells and ~1.7 media ml/min per g in the U87-Δ45-TGL cells. The bioluminescence was ~2.8 × 10<sup>3</sup> relative bioluminescence unit (RLU) in the U87-NES-TGL cells and ~4.9 × 10<sup>3</sup> RLU in the U87-Δ45-TGL. The wild-type U87 cells were used as a control. Their measured fluorescence, influx rate, and bioluminescence were negligible.</p></div><div id="TGL.Animal_Studies"><h2 id="_TGL_Animal_Studies_">Animal Studies</h2><div id="TGL.Rodents"><h3>Rodents</h3><p>[<a href="/sites/entrez?Db=pubmed&Cmd=DetailsSearch&Term=(%20HSV1-tk%20+%20GFP%20+%20luciferase)%20+AND++rodentia" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">PubMed</a>]</p><p>Ponomarev et al. used three imaging modalities to examine the expression of the triple reporter proteins <i>in vivo</i> (<a class="bk_pop" href="#TGL.REF.4">4</a>). <i>Nu</i>/<i>nu</i> mice (20–25 g) were implanted subcutaneously with U87-NES-TGL cells on the right shoulder, with U87-Δ45-TGL cells on the left shoulder, and with wild-type U87 cells on the left flank. The tumors grew to an average size of 5.4 ± 2.1 mm and were imaged with whole-body fluorescence, bioluminescence, and gamma camera imaging. For bioluminescence imaging, mice were injected intravenously with 100 μl of D-luciferin solution at a dose of 150 mg/kg, and the images were acquired for 0.5–10 s. For the gamma camera imaging, mice received an intraperitoneal injection of 0.9% NaI solution (1 ml) one day before imaging to block the thyroid uptake of radioactive iodide. Then the mice were injected intravenously with [<sup>131</sup>I]-FIAU at a dose of 7.4 MBq per animal. The gamma camera images were collected 24 h after [<sup>131</sup>I]-FIAU injection to allow sufficient clearance of body background radioactivity. The GFP fluorescence was collected with a charge-coupled device (CCD) camera. The average level of gene expression in the implanted U87-NES-TGL tumor was detected as 707 ± 36 in fluorescence, 486 ± 47 × 10<sup>6</sup> photons/cm<sup>2</sup> per sr in bioluminescence, and 0.47 ± 0.08% injected dose (ID)/g in [<sup>131</sup>I]-FIAU accumulation. The average level of gene expression in the implanted U87-Δ45-TGL tumor was detected as 740 ± 47 in fluorescence, 508 ± 36 × 10<sup>6</sup> photons/cm<sup>2</sup> per sr in bioluminescence, and 0.86 ± 0.06% ID/g in [<sup>131</sup>I]-FIAU accumulation. As a control, the average level of gene expression in the implanted wild-type U87 cells was found at background level in fluorescence or bioluminescence imaging and at 0.03 ± 0.01% ID/g in [<sup>131</sup>I]-FIAU accumulation (the background level). After completion of the triple modality imaging, the mice were euthanized and the tissues were harvested for <i>in vitro</i> fluorescence imaging. Bright green fluorescent areas were found in the tumor tissues, demonstrating the presence of TGL labeling.</p></div><div id="TGL.Other_NonPrimate_Mam"><h3>Other Non-Primate Mammals</h3><p>[<a href="/entrez/query.fcgi?cmd=PureSearch&db=pubmed&details_term=%22%20SUBSTANCENAME%22%5BSubstance%20Name%5D%20AND%20%28dog%20OR%20rabbit%20OR%20pig%20OR%20sheep%29" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">PubMed</a>]</p><p>No publication is currently available.</p></div><div id="TGL.NonHuman_Primates"><h3>Non-Human Primates</h3><p>[<a href="/sites/entrez?Db=pubmed&Cmd=DetailsSearch&Term=(%20HSV1-tk%20+%20GFP%20+%20luciferase)%20+AND+(primate+non+human)" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">PubMed</a>]</p><p>No publication is currently available.</p></div></div><div id="TGL.Human_Studies"><h2 id="_TGL_Human_Studies_">Human Studies</h2><p>[<a href="/sites/entrez?Db=pubmed&Cmd=DetailsSearch&Term=(%20HSV1-tk%20+%20GFP%20+%20luciferase)%20+AND+human" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">PubMed</a>]</p><p>No publication is currently available.</p></div><div id="TGL.references"><h2 id="_TGL_references_">References</h2><dl class="temp-labeled-list"><dt>1.</dt><dd><div class="bk_ref" id="TGL.REF.1">Massoud T.F. , Singh A. , Gambhir S.S. Noninvasive molecular neuroimaging using reporter genes: part I, principles revisited. <span><span class="ref-journal">AJNR Am J Neuroradiol. </span>2008;<span class="ref-vol">
|
||
<strong>29</strong>
|
||
</span>(2):229–34.</span> [<a href="/pmc/articles/PMC8118983/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC8118983</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/18024575" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 18024575</span></a>]</div></dd><dt>2.</dt><dd><div class="bk_ref" id="TGL.REF.2">Gross S. , Piwnica-Worms D. Spying on cancer: molecular imaging in vivo with genetically encoded reporters. <span><span class="ref-journal">Cancer Cell. </span>2005;<span class="ref-vol">
|
||
<strong>7</strong>
|
||
</span>(1):5–15.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/15652745" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 15652745</span></a>]</div></dd><dt>3.</dt><dd><div class="bk_ref" id="TGL.REF.3">Serganova I. , Ponomarev V. , Blasberg R. Human reporter genes: potential use in clinical studies. <span><span class="ref-journal">Nucl Med Biol. </span>2007;<span class="ref-vol">
|
||
<strong>34</strong>
|
||
</span>(7):791–807.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/17921031" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 17921031</span></a>]</div></dd><dt>4.</dt><dd><div class="bk_ref" id="TGL.REF.4">Ponomarev V. , Doubrovin M. , Serganova I. , Vider J. , Shavrin A. , Beresten T. , Ivanova A. , Ageyeva L. , Tourkova V. , Balatoni J. , Bornmann W. , Blasberg R. , Gelovani Tjuvajev J. A novel triple-modality reporter gene for whole-body fluorescent, bioluminescent, and nuclear noninvasive imaging. <span><span class="ref-journal">Eur J Nucl Med Mol Imaging. </span>2004;<span class="ref-vol">
|
||
<strong>31</strong>
|
||
</span>(5):740–51.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/15014901" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 15014901</span></a>]</div></dd><dt>5.</dt><dd><div class="bk_ref" id="TGL.REF.5">Wurth C. , Thomas R.M. , Folkers G. , Scapozza L. Folding and self-assembly of herpes simplex virus type 1 thymidine kinase. <span><span class="ref-journal">J Mol Biol. </span>2001;<span class="ref-vol">
|
||
<strong>313</strong>
|
||
</span>(3):657–70.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/11676546" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 11676546</span></a>]</div></dd><dt>6.</dt><dd><div class="bk_ref" id="TGL.REF.6">Ponomarev V. , Doubrovin M. , Serganova I. , Beresten T. , Vider J. , Shavrin A. , Ageyeva L. , Balatoni J. , Blasberg R. , Tjuvajev J.G. Cytoplasmically retargeted HSV1-tk/GFP reporter gene mutants for optimization of noninvasive molecular-genetic imaging. <span><span class="ref-journal">Neoplasia. </span>2003;<span class="ref-vol">
|
||
<strong>5</strong>
|
||
</span>(3):245–54.</span> [<a href="/pmc/articles/PMC1502405/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC1502405</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/12869307" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 12869307</span></a>]</div></dd><dt>7.</dt><dd><div class="bk_ref" id="TGL.REF.7">Kussmann-Gerber S. , Kuonen O. , Folkers G. , Pilger B.D. , Scapozza L. Drug resistance of herpes simplex virus type 1--structural considerations at the molecular level of the thymidine kinase. <span><span class="ref-journal">Eur J Biochem. </span>1998;<span class="ref-vol">
|
||
<strong>255</strong>
|
||
</span>(2):472–81.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/9716390" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 9716390</span></a>]</div></dd><dt>8.</dt><dd><div class="bk_ref" id="TGL.REF.8">Chalfie M. , Tu Y. , Euskirchen G. , Ward W.W. , Prasher D.C. Green fluorescent protein as a marker for gene expression. <span><span class="ref-journal">Science. </span>1994;<span class="ref-vol">
|
||
<strong>263</strong>
|
||
</span>(5148):802–5.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/8303295" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 8303295</span></a>]</div></dd><dt>9.</dt><dd><div class="bk_ref" id="TGL.REF.9">Conti E. , Franks N.P. , Brick P. Crystal structure of firefly luciferase throws light on a superfamily of adenylate-forming enzymes. <span><span class="ref-journal">Structure. </span>1996;<span class="ref-vol">
|
||
<strong>4</strong>
|
||
</span>(3):287–98.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/8805533" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 8805533</span></a>]</div></dd><dt>10.</dt><dd><div class="bk_ref" id="TGL.REF.10">Paulmurugan R. , Gambhir S.S. Firefly luciferase enzyme fragment complementation for imaging in cells and living animals. <span><span class="ref-journal">Anal Chem. </span>2005;<span class="ref-vol">
|
||
<strong>77</strong>
|
||
</span>(5):1295–302.</span> [<a href="/pmc/articles/PMC4154832/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC4154832</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/15732910" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 15732910</span></a>]</div></dd><dt>11.</dt><dd><div class="bk_ref" id="TGL.REF.11">Jacobs A. , Dubrovin M. , Hewett J. , Sena-Esteves M. , Tan C.W. , Slack M. , Sadelain M. , Breakefield X.O. , Tjuvajev J.G. Functional coexpression of HSV-1 thymidine kinase and green fluorescent protein: implications for noninvasive imaging of transgene expression. <span><span class="ref-journal">Neoplasia. </span>1999;<span class="ref-vol">
|
||
<strong>1</strong>
|
||
</span>(2):154–61.</span> [<a href="/pmc/articles/PMC1508134/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC1508134</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/10933050" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 10933050</span></a>]</div></dd></dl></div><div id="bk_toc_contnr"></div></div></div>
|
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<div class="post-content"><div><div class="half_rhythm"><a href="/books/about/copyright/">Copyright Notice</a></div><div class="small"><span class="label">Bookshelf ID: NBK23131</span><span class="label">PMID: <a href="https://pubmed.ncbi.nlm.nih.gov/20641334" title="PubMed record of this page" ref="pagearea=meta&targetsite=entrez&targetcat=link&targettype=pubmed">20641334</a></span></div><div style="margin-top:2em" class="bk_noprnt"><a class="bk_cntns" href="/books/n/micad/">Contents</a><div class="pagination bk_noprnt"><a class="active page_link prev" href="/books/n/micad/sosgreenfluorescencepro2/" title="Previous page in this title">< Prev</a><a class="active page_link next" href="/books/n/micad/soshilytefluor647/" title="Next page in this title">Next ></a></div></div></div></div>
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<div xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance"></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>Views</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="PDF_download" id="Shutter"></a></div><div class="portlet_content"><ul xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="simple-list"><li><a href="/books/NBK23131/?report=reader">PubReader</a></li><li><a href="/books/NBK23131/?report=printable">Print View</a></li><li><a data-jig="ncbidialog" href="#_ncbi_dlg_citbx_NBK23131" data-jigconfig="width:400,modal:true">Cite this Page</a><div id="_ncbi_dlg_citbx_NBK23131" style="display:none" title="Cite this Page"><div class="bk_tt">Zhang H. HSV1-TK/GFP/Fluc. 2008 Oct 21 [Updated 2008 Dec 1]. In: Molecular Imaging and Contrast Agent Database (MICAD) [Internet]. Bethesda (MD): National Center for Biotechnology Information (US); 2004-2013. <span class="bk_cite_avail"></span></div></div></li><li><a href="/books/NBK23131/pdf/Bookshelf_NBK23131.pdf">PDF version of this page</a> (136K)</li><li><a href="/books/n/micad/toc/bin/micad.csv">MICAD summary (CSV file)</a></li></ul></div></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>In this page</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="page-toc" id="Shutter"></a></div><div class="portlet_content"><ul xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="simple-list"><li><a href="#TGL.Background" ref="log$=inpage&link_id=inpage">Background</a></li><li><a href="#TGL.Synthesis" ref="log$=inpage&link_id=inpage">Synthesis</a></li><li><a href="#TGL.In_Vitro_Studies_Tes" ref="log$=inpage&link_id=inpage"><i>In Vitro</i> Studies: Testing in Cells and Tissues</a></li><li><a href="#TGL.Animal_Studies" ref="log$=inpage&link_id=inpage">Animal Studies</a></li><li><a href="#TGL.Human_Studies" ref="log$=inpage&link_id=inpage">Human Studies</a></li><li><a href="#TGL.references" ref="log$=inpage&link_id=inpage">References</a></li></ul></div></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>Search MICAD</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="source-application" id="Shutter"></a></div><div class="portlet_content"><form xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" name="frmSearch" method="get" action="/books/NBK5330/" id="frmSearch"><script type="text/javascript" src="/corehtml/pmc//js/bookshelf/micad.js">/**/</script><label class="offscreen_noflow" for="txtfield">Search term</label><input id="txtfield" type="text" name="f1_term" size="22" onKeyPress="KeyPress('micad',event,'/books/NBK5330/','')" /><button name="f1_search" type="submit">Go</button><button onclick="this.form.reset();" type="reset">Clear</button><p><b>Limit my Search:</b></p><div class="clearfix"><label for="detection">Method of detection:</label><div class="right"><select name="detection" id="detection" style="width:200px"><option value="" selected="selected">Any</option><option value="(MRI OR "Magnetic resonance imaging" OR MRS)">MRI</option><option value="Multimodal">Multimodal imaging</option><option value="Optical">Optical imaging</option><option value="PET">PET</option><option value="Photoacoustic">Photoacoustic imaging</option><option value="(SPECT OR planar)">SPECT</option><option value="Ultrasound">Ultrasound</option><option value="(x-ray OR ct)">X-ray, CT</option></select></div></div><div class="clearfix"><label for="signal">Source of signal/contrast:</label><div class="right"><select name="signal" id="signal" style="width:200px"><option value="" selected="selected">Any</option><optgroup label="MRI agents"><option value="(Copper OR Cu)">Copper</option><option value="(Europium OR Eu3+)">Europium</option><option value="(Fluorine OR 19F)">Fluorine</option><option value="(Gadolinium OR Gd3+)">Gadolinium</option><option value=""Hyperpolarized 13C"">Hyperpolarized 13C</option><option value=""Iron oxide"">Iron oxide</option><option value=""Nitroxide radicals"">Nitroxide radicals</option><option value="(Oxygen OR 17O)">Oxygen</option><option value="Thulium">Thulium</option></optgroup><optgroup label="Multimodal agents"><option value="((Gadolinium OR Gd3+) AND Optical)">Gadolinium and optical</option><option value="((Gadolinium OR Gd3+) AND (Gold OR Au))">Gadolinium and Gold</option><option value="("Iron oxide" AND (64Cu OR 124I OR 111In))">Iron oxide and
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value="("Nitrogen 13" OR 13N)">Nitrogen-13</option><option value="("Yttrium 86" OR 86Y)">Yttrium-86</option><option value="("Zirconium 89" OR 89Zr)">Zirconium-89</option></optgroup><optgroup label="Photoacoustic agents"><option value="(Gold OR Au)">Gold</option><option value="("Indocyanine green" OR ICG)">Indocyanine green</option></optgroup><optgroup label="SPECT radionuclides"><option value="(Gallium-67 OR 67Ga)">Gallium-67</option><option value="("Indium 111" OR 111In)">Indium-111</option><option value="("Iodine 123" OR "Iodine 125" OR "Iodine 131" OR 123I OR 125I OR 131I)">Iodine-123, 125, 131</option><option value="("Lutetium 177" OR 177Lu)">Lutetium-177</option><option value="(Rhenium OR 186Re OR 188Re)">Rhenium</option><option value="("Technetium 99m" OR 99mTc)">Technetium-99m</option><option value="("Tellurium 125m" OR 125mTe)">Tellurium-125m</option></optgroup><optgroup label="Ultrasound agents"><option value="Microbubbles">Microbubbles</option><option value="Nanobubbles">Nanobubbles</option></optgroup><optgroup label="X-ray and CT agents"><option value="(Bismuth OR Bi)">Bismuth</option><option value="(Gold OR Au)">Gold</option><option value="Iodine">Iodine</option></optgroup></select></div></div><div class="clearfix"><label for="agent">Agent Category:</label><div class="right"><select name="agent" id="agent" style="width:200px"><option value="" selected="selected">Any</option><option value="(antibody OR trastuzumab OR immunoglobulin)">Antibodies</option><option value="(bacteria OR bacteriophage OR coli)">Bacteria</option><option value="cell">Cells</option><option value="(compound OR "amino acid" OR "folic acid" OR "cage molecule" OR carbohydrate OR copolymers OR polymer OR "small molecule" OR macromolecule OR triiodobenzoate OR estradiol OR glycosaminoglycan)">Compounds</option><option value="ligand">Ligands</option><option value="(lipid OR liposome OR liposomes">Lipids</option><option value="metal">Metal</option><option value="(nanoparticle OR nanoparticles OR nanotubes OR "iron oxide")">Nanoparticles</option><option value="(siRNA OR "nucleic acid" OR oligonucleotide)">Nucleic acids</option><option value="peptide">Peptides</option><option value="polyeptide">Polyeptides</option><option value="(protein OR albumin OR chemokin OR immunoprotein OR luciferase OR albumin)">Proteins</option><option value="(virus OR adenovirus)">Viruses</option></select></div></div><div class="clearfix"><label for="target">Target Category:</label><div class="right"><select name="target" id="target" style="width:200px"><option value="" selected="selected">Any</option><option value="acceptor">Acceptors</option><option value=""adhesion molecule"">Adhesion molecules</option><option value="(antigen OR antibody-antigen)">Antigens</option><option value="(enzyme OR enzymes OR enzyme-substrate)">Enzymes</option><option value="(lipids OR lipophilic cation)">Lipids</option><option value="(receptor OR receptors OR receptor-ligand OR receptor-antibody OR antibody-receptor)">Receptors</option><option value="transporter">Transporters</option><option value="non-targeted">Non-targeted</option><option value=""nucleic acid"">Nucleic acids</option><option value="(non-targeted OR "unknown binding site")">Others</option></select></div></div><div><input id="__micad_btn_1" type="radio" name="stage" value="vitro" /><label for="__micad_btn_1"><i>In vitro</i></label><input id="__micad_btn_2" type="radio" name="stage" value="rodents" /><label for="__micad_btn_2">Rodents</label><input id="__micad_btn_3" type="radio" name="stage" value="mammals" /><label for="__micad_btn_3">Non-primate non-rodent mammals</label><br /><input id="__micad_btn_4" type="radio" name="stage" value="primates" /><label for="__micad_btn_4">Non-human primates</label><input id="__micad_btn_5" type="radio" name="stage" value="humans" /><label for="__micad_btn_5">Humans</label><input id="__micad_btn_6" type="radio" name="stage" value="any" checked="checked" /><label for="__micad_btn_6">Any</label></div></form><form xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" name="frmGo" method="get" action="javascript:alert('frmGo:_@action_was_not_set')" id="frmGo"><input name="term" value="." type="hidden" /></form></div></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>Related information</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="discovery_db_links" id="Shutter"></a></div><div class="portlet_content"><ul><li class="brieflinkpopper"><a class="brieflinkpopperctrl" href="/books/?Db=pmc&DbFrom=books&Cmd=Link&LinkName=books_pmc_refs&IdsFromResult=1505230" ref="log$=recordlinks">PMC</a><div class="brieflinkpop offscreen_noflow">PubMed Central citations</div></li><li class="brieflinkpopper"><a class="brieflinkpopperctrl" href="/books/?Db=pubmed&DbFrom=books&Cmd=Link&LinkName=books_pubmed_refs&IdsFromResult=1505230" ref="log$=recordlinks">PubMed</a><div class="brieflinkpop offscreen_noflow">Links to PubMed</div></li></ul></div></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>Similar articles in PubMed</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="PBooksDiscovery_RA" id="Shutter"></a></div><div class="portlet_content"><ul><li class="brieflinkpopper two_line"><a class="brieflinkpopperctrl" href="/pubmed/20641191" ref="ordinalpos=1&linkpos=1&log$=relatedreviews&logdbfrom=pubmed"><span xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="invert">Review</span> Ad5-(PSE-BC)-(GAL4-(VP16)(2))-(GAL4)(5)-sr39tk.</a><span class="source">[Molecular Imaging and Contrast...]</span><div class="brieflinkpop offscreen_noflow"><span 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class="brieflinkpop offscreen_noflow"><span xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="invert">Review</span> 2'-Fluoro-2'-deoxy-5'-[(123/124/125/131)I]iodo-1β-d-arabinofuranosyluracil.<div class="brieflinkpopdesc"><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="author">Leung K. </em><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="cit">Molecular Imaging and Contrast Agent Database (MICAD). 2004</em></div></div></li><li class="brieflinkpopper two_line"><a class="brieflinkpopperctrl" href="/pubmed/15014901" ref="ordinalpos=1&linkpos=3&log$=relatedarticles&logdbfrom=pubmed">A novel triple-modality reporter gene for whole-body fluorescent, bioluminescent, and nuclear noninvasive imaging.</a><span class="source">[Eur J Nucl Med Mol Imaging. 2004]</span><div class="brieflinkpop offscreen_noflow">A 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