{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T18:40:41Z","timestamp":1792694441597,"version":"4.5.0"},"reference-count":37,"publisher":"MyJove Corporation","issue":"83","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JoVE"],"DOI":"10.3791\/51151","type":"journal-article","created":{"date-parts":[[2014,1,17]],"date-time":"2014-01-17T18:59:53Z","timestamp":1389985193000},"source":"Crossref","is-referenced-by-count":6,"title":["Methodology for the Efficient Generation of Fluorescently Tagged Vaccinia Virus Proteins"],"prefix":"10.3791","author":[{"given":"N. Bishara","family":"Marzook","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dean J.","family":"Procter","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Helena","family":"Lynn","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yui","family":"Yamamoto","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jacquelyn","family":"Horsington","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Timothy P.","family":"Newsome","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"2098","published-online":{"date-parts":[[2014,1,17]]},"reference":[{"key":"ref1","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1111\/j.1574-6976.2000.tb00536.x","type":"journal-article","article-title":"Adventures with poxviruses of vertebrates","volume":"24","author":"Fenner","year":"2000","unstructured":"Fenner, F. Adventures with poxviruses of vertebrates. FEMS Microbiol. Rev. 24, 123-133 (2000).","journal-title":"FEMS Microbiol. Rev"},{"key":"ref2","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1016\/0042-6822(90)90294-2","type":"journal-article","article-title":"The Complete DNA-Sequence of Vaccinia Virus","volume":"179","author":"Goebel","year":"1990","unstructured":"Goebel, S. J. et al. The Complete DNA-Sequence of Vaccinia Virus. Virology. 179, 247-266 (1990).","journal-title":"Virology"},{"key":"ref3","doi-asserted-by":"crossref","first-page":"1349","DOI":"10.1099\/0022-1317-72-6-1349","type":"journal-article","article-title":"Nucleotide-sequence of 42 kbp of vaccinia virus-strain WR from near the right inverted terminal repeat","volume":"72","author":"Smith","year":"1991","unstructured":"Smith, G. L., Chan, Y. S. & Howard, S. T. Nucleotide-sequence of 42 kbp of vaccinia virus-strain WR from near the right inverted terminal repeat J. Gen. Virol. 72, 1349-1376 (1991).","journal-title":"J. Gen. Virol"},{"key":"ref4","doi-asserted-by":"crossref","first-page":"472","DOI":"10.1016\/j.tim.2008.07.009","type":"journal-article","article-title":"Vaccinia virus morphogenesis and dissemination","volume":"16","author":"Roberts","year":"2008","unstructured":"Roberts, K. L. & Smith, G. L. Vaccinia virus morphogenesis and dissemination. Trends Microbiol. 16, 472-479 (2008).","journal-title":"Trends Microbiol"},{"key":"ref5","doi-asserted-by":"crossref","first-page":"4236","DOI":"10.1128\/JVI.02255-08","type":"journal-article","article-title":"The 3 '-to-5 ' Exonuclease Activity of Vaccinia Virus DNA Polymerase Is Essential and Plays a Role in Promoting Virus Genetic Recombination","volume":"83","author":"Gammon","year":"2009","unstructured":"Gammon, D. B. & Evans, D. H. The 3 '-to-5 ' Exonuclease Activity of Vaccinia Virus DNA Polymerase Is Essential and Plays a Role in Promoting Virus Genetic Recombination. J. Virol. 83, 4236-4250 (2009).","journal-title":"J. Virol"},{"key":"ref6","doi-asserted-by":"crossref","first-page":"6923","DOI":"10.1128\/JVI.75.15.6923-6932.2001","type":"journal-article","article-title":"Effects of DNA structure and homology length on vaccinia virus recombination","volume":"75","author":"Yao","year":"2001","unstructured":"Yao, X. D. & Evans, D. H. Effects of DNA structure and homology length on vaccinia virus recombination. J. Virol. 75, 6923-6932 (2001).","journal-title":"J. Virol"},{"key":"ref7","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1385\/1-59259-789-0:205","type":"journal-article","volume":"16","author":"Ward","year":"2004","unstructured":"Ward, B. in Vaccinia Virus and Poxvirology Vol. 269 Methods in Molecular Biology. ed StuartN Isaacs. Humana Press, Ch. 16, 205-218 (2004).","journal-title":"Ch. 16 Vaccinia Virus and Poxvirology Vol. 269 Methods in Molecular Biology"},{"key":"ref8","first-page":"21","type":"journal-article","article-title":"Infectious Poxvirus Vectors Have Capacity for at Least 25,000","volume":"25","author":"Smith","year":"1983","unstructured":"Smith, G. L. & Moss, B. Infectious Poxvirus Vectors Have Capacity for at Least 25,000 Base-Pairs of Foreign DNA. Gene. 25, 21-28 (1983).","journal-title":"Base-Pairs of Foreign DNA. Gene"},{"key":"ref9","doi-asserted-by":"crossref","first-page":"269","DOI":"10.1083\/jcb.200412169","type":"journal-article","article-title":"Deep-etch EM reveals that the early poxvirus envelope is a single membrane bilayer stabilized by a geodetic &#34;honeycomb&#34; surface coat","volume":"169","author":"Heuser","year":"2005","unstructured":"Heuser, J. Deep-etch EM reveals that the early poxvirus envelope is a single membrane bilayer stabilized by a geodetic \"honeycomb\" surface coat. J. Cell Biol. 169, 269-283 (2005).","journal-title":"J. Cell Biol"},{"key":"ref10","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1016\/j.coviro.2011.11.007","type":"journal-article","article-title":"Poxvirus host cell entry","volume":"2","author":"Schmidt","year":"2012","unstructured":"Schmidt, F. I., Bleck, C. K. E. & Mercer, J. Poxvirus host cell entry. Curr. Opin. Virol. 2, 20-27 (2012).","journal-title":"Curr. Opin. Virol"},{"key":"ref11","doi-asserted-by":"crossref","first-page":"4755","DOI":"10.1128\/JVI.79.8.4755-4763.2005","type":"journal-article","article-title":"Visualization and characterization of the intracellular movement of vaccinia virus intracellular mature virions","volume":"79","author":"Ward","year":"2005","unstructured":"Ward, B. M. Visualization and characterization of the intracellular movement of vaccinia virus intracellular mature virions. J. Virol. 79, 4755-4763 (2005).","journal-title":"J. Virol"},{"key":"ref12","doi-asserted-by":"crossref","first-page":"2443","DOI":"10.1099\/vir.0.19271-0","type":"journal-article","article-title":"Vaccinia virus cores are transported on microtubules","volume":"84","author":"Carter","year":"2003","unstructured":"Carter, G. C. et al. Vaccinia virus cores are transported on microtubules. J. Gen. Virol. 84, 2443-2458 (2003).","journal-title":"J. Gen. Virol"},{"key":"ref13","doi-asserted-by":"crossref","first-page":"4802","DOI":"10.1128\/JVI.75.10.4802-4813.2001","type":"journal-article","article-title":"Visualization of intracellular movement of vaccinia virus virions containing a green fluorescent protein-B5R membrane protein chimera","volume":"75","author":"Ward","year":"2001","unstructured":"Ward, B. M. & Moss, B. Visualization of intracellular movement of vaccinia virus virions containing a green fluorescent protein-B5R membrane protein chimera. J. Virol. 75, 4802-4813 (2001).","journal-title":"J. Virol"},{"key":"ref14","doi-asserted-by":"crossref","first-page":"997","DOI":"10.1128\/jvi.63.2.997-1001.1989","type":"journal-article","article-title":"Plaque size phenotype as a selectable marker to generate vaccinia virus recombinants","volume":"63","author":"Rodriguez","year":"1989","unstructured":"Rodriguez, J. F. & Esteban, M. Plaque size phenotype as a selectable marker to generate vaccinia virus recombinants. J. Virol. 63, 997-1001 (1989).","journal-title":"J. Virol"},{"key":"ref15","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1016\/0378-1119(95)00149-Z","type":"journal-article","article-title":"Selection of recombinant vaccinia viruses on the basis of plaque-formation","volume":"158","author":"Blasco","year":"1995","unstructured":"Blasco, R. & Moss, B. Selection of recombinant vaccinia viruses on the basis of plaque-formation. Gene. 158, 157-162 (1995).","journal-title":"Gene"},{"key":"ref16","doi-asserted-by":"crossref","first-page":"7415","DOI":"10.1073\/pnas.79.23.7415","type":"journal-article","article-title":"Vaccinia virus - a selectable eukaryotic cloning and expression vector","volume":"79","author":"Mackett","year":"1982","unstructured":"Mackett, M., Smith, G. L. & Moss, B. Vaccinia virus - a selectable eukaryotic cloning and expression vector. Proc. Natl. Acad. Sci. U.S.A. 79, 7415-7419 (1982).","journal-title":"Proc. Natl. Acad. Sci. U.S.A"},{"key":"ref17","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1016\/0378-1119(86)90063-6","type":"journal-article","article-title":"Vaccinia virus vectors utilizing the beta-galactosidase assay for rapid selection of recombinant viruses and measurement of gene-expression","volume":"47","author":"Panicali","year":"1986","unstructured":"Panicali, D., Grzelecki, A. & Huang, C. Vaccinia virus vectors utilizing the beta-galactosidase assay for rapid selection of recombinant viruses and measurement of gene-expression Gene. 47, 193-199 (1986).","journal-title":"Gene"},{"key":"ref18","doi-asserted-by":"crossref","first-page":"1849","DOI":"10.1128\/jvi.62.6.1849-1854.1988","type":"journal-article","article-title":"Escherichia-coli gpt gene provides dominant selection for vaccinia virus open reading frame expression vectors","volume":"62","author":"Falkner","year":"1988","unstructured":"Falkner, F. G. & Moss, B. Escherichia-coli gpt gene provides dominant selection for vaccinia virus open reading frame expression vectors. J. Virol. 62, 1849-1854 (1988).","journal-title":"J. Virol"},{"key":"ref19","first-page":"188","type":"journal-article","article-title":"Selection of recombinant vaccinia viruses (Tian-Tan strain) expressing hepatitis-B virus surface-antigen by using beta-galactosidase as a marker","volume":"33","author":"Liu","year":"1990","unstructured":"Liu, G. Q. et al. Selection of recombinant vaccinia viruses (Tian-Tan strain) expressing hepatitis-B virus surface-antigen by using beta-galactosidase as a marker. Sci. China Ser. B-Chem. 33, 188-197 (1990).","journal-title":"Sci. China Ser. B-Chem"},{"key":"ref20","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1016\/S0022-1759(98)00156-2","type":"journal-article","article-title":"Green fluorescent protein expressed by a recombinant vaccinia virus permits early detection of infected cells by flow cytometry","volume":"220","author":"Dom\u00ednguez","year":"1998","unstructured":"Dom\u00ednguez, J., Lorenzo, M. D. M. & Blasco, R. Green fluorescent protein expressed by a recombinant vaccinia virus permits early detection of infected cells by flow cytometry. J. Immunol. Methods. 220, 115-121 (1998).","journal-title":"J. Immunol. Methods"},{"key":"ref21","doi-asserted-by":"crossref","first-page":"3108","DOI":"10.1128\/jvi.64.6.3108-3111.1990","type":"journal-article","article-title":"Transient dominant selection of recombinant vaccinia viruses","volume":"64","author":"Falkner","year":"1990","unstructured":"Falkner, F. G. & Moss, B. Transient dominant selection of recombinant vaccinia viruses. J. Virol. 64, 3108-3111 (1990).","journal-title":"J. Virol"},{"key":"ref22","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0008506","type":"journal-article","article-title":"F11-Mediated Inhibition of RhoA Signalling Enhances the Spread of Vaccinia Virus In Vitro and In Vivo in an Intranasal Mouse Model of Infection","volume":"4","author":"Cordeiro","year":"2009","unstructured":"Cordeiro, J. V. et al. F11-Mediated Inhibition of RhoA Signalling Enhances the Spread of Vaccinia Virus In Vitro and In Vivo in an Intranasal Mouse Model of Infection. Plos One. 4, (2009).","journal-title":"Plos One"},{"key":"ref23","doi-asserted-by":"crossref","first-page":"7485","DOI":"10.1128\/jvi.70.11.7485-7497.1996","type":"journal-article","article-title":"Identification of the major membrane and core proteins of vaccinia virus by two-dimensional electrophoresis","volume":"70","author":"Jensen","year":"1996","unstructured":"Jensen, O. N. et al. Identification of the major membrane and core proteins of vaccinia virus by two-dimensional electrophoresis. J. Virol. 70, 7485-7497 (1996).","journal-title":"J. Virol"},{"key":"ref24","doi-asserted-by":"crossref","first-page":"757","DOI":"10.1128\/jvi.58.3.757-764.1986","type":"journal-article","article-title":"Localization and Fine-Structure of a Vaccinia Virus Gene Encoding an Envelope Antigen","volume":"58","author":"Hirt","year":"1986","unstructured":"Hirt, P., Hiller, G. & Wittek, R. Localization and Fine-Structure of a Vaccinia Virus Gene Encoding an Envelope Antigen. J. Virol. 58, 757-764, (1986).","journal-title":"J. Virol"},{"key":"ref25","first-page":"445","type":"journal-article","article-title":"An improved cyan fluorescent protein variant useful for","volume":"22","author":"Rizzo","year":"2004","unstructured":"Rizzo, M. A., Springer, G. H., Granada, B. & Piston, D. W. An improved cyan fluorescent protein variant useful for FRET. Nat. Biotechnol. 22, 445-449 (2004)."},{"key":"ref26","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pbio.1001041","type":"journal-article","article-title":"A Genetically Encoded Tag for Correlated Light and Electron Microscopy of Intact Cells, Tissues, and Organisms","volume":"9","author":"Shu","year":"2011","unstructured":"Shu, X. K. et al. A Genetically Encoded Tag for Correlated Light and Electron Microscopy of Intact Cells, Tissues, and Organisms. Plos Biol. 9 (2011).","journal-title":"Plos Biol"},{"key":"ref27","doi-asserted-by":"crossref","first-page":"295","DOI":"10.1016\/j.jviromet.2010.11.003","type":"journal-article","article-title":"Engineering recombinant poxviruses using a compact GFP-blasticidin resistance fusion gene for selection","volume":"171","author":"Wong","year":"2011","unstructured":"Wong, Y. C., Lin, L. C. W., Melo-Silva, C. R., Smith, S. A. & Tscharke, D. C. Engineering recombinant poxviruses using a compact GFP-blasticidin resistance fusion gene for selection. J. Virol. Methods. 171, 295-298 (2011).","journal-title":"J. Virol. Methods"},{"key":"ref28","first-page":"173","type":"journal-article","volume":"62","author":"Broder","year":"1997","unstructured":"Broder, C. C. & Earl, P. L. Vol. 62 173-197 (1997)."},{"key":"ref29","doi-asserted-by":"crossref","first-page":"5910","DOI":"10.1128\/jvi.65.11.5910-5920.1991","type":"journal-article","article-title":"Extracellular Vaccinia virus formation and cell-to-cell virus transmission are prevented by deletion of the gene encoding the 37,000-dalton outer envelope protein","volume":"65","author":"Blasco","year":"1991","unstructured":"Blasco, R. & Moss, B. Extracellular Vaccinia virus formation and cell-to-cell virus transmission are prevented by deletion of the gene encoding the 37,000-dalton outer envelope protein. J. Virol. 65, 5910-5920 (1991).","journal-title":"J. Virol"},{"key":"ref30","first-page":"155","type":"journal-article","article-title":"Ch. Navigating the subcellular space: Lessons from vaccinia virus","author":"Newsome","year":"2011","unstructured":"Newsome, T. P., Marty, A. J., Lynn, H., Procter, D.J. . in Viral Transport, Assembly and Egress. ed R.J. Diefenbach, Cunningham, A.L.. Research Signpost Ch. Navigating the subcellular space: Lessons from vaccinia virus, 155-177 (2011).","journal-title":"Viral Transport, Assembly and Egress"},{"key":"ref31","doi-asserted-by":"crossref","first-page":"12886","DOI":"10.1128\/JVI.01395-10","type":"journal-article","article-title":"Interaction of Poxvirus Intracellular Mature Virion Proteins with the TPR Domain of Kinesin Light Chain in Live Infected Cells Revealed by Two-Photon-Induced Fluorescence Resonance Energy Transfer Fluorescence Lifetime Imaging Microscopy","volume":"84","author":"Jeshtadi","year":"2010","unstructured":"Jeshtadi, A. et al. Interaction of Poxvirus Intracellular Mature Virion Proteins with the TPR Domain of Kinesin Light Chain in Live Infected Cells Revealed by Two-Photon-Induced Fluorescence Resonance Energy Transfer Fluorescence Lifetime Imaging Microscopy. J. Virol. 84, 12886-12894 (2010).","journal-title":"J. Virol"},{"key":"ref32","doi-asserted-by":"crossref","DOI":"10.1038\/nature07773","type":"journal-article","article-title":"The rate of N-WASP exchange limits the extent of ARP2\/3-complex-dependent actin-based motility","volume":"458","author":"Weisswange","year":"2009","unstructured":"Weisswange, I., Newsome, T. P., Schleich, S. & Way, M. The rate of N-WASP exchange limits the extent of ARP2\/3-complex-dependent actin-based motility. Nature. 458, 87-U86 (2009).","journal-title":"Nature"},{"key":"ref33","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1016\/j.antiviral.2011.04.014","type":"journal-article","article-title":"Development of Vaccinia reporter viruses for rapid, high content analysis of viral function at all stages of gene expression","volume":"91","author":"Dower","year":"2011","unstructured":"Dower, K., Rubins, K. H., Hensley, L. E. & Connor, J. H. Development of Vaccinia reporter viruses for rapid, high content analysis of viral function at all stages of gene expression. Antiviral Res. 91, 72-80 (2011).","journal-title":"Antiviral Res"},{"key":"ref34","doi-asserted-by":"crossref","first-page":"252","DOI":"10.2174\/1874070700802010252","type":"journal-article","article-title":"Engineering oncolytic Vaccinia viruses for non-invasive optical imaging of tumors","volume":"2","author":"D\u00e9nes","year":"2008","unstructured":"D\u00e9nes, B., Fodor, N., Obenaus, A. & F., I. Engineering oncolytic Vaccinia viruses for non-invasive optical imaging of tumors. Open Biotechnol. J. 2, 252-261 (2008).","journal-title":"Open Biotechnol. J"},{"key":"ref35","doi-asserted-by":"crossref","first-page":"346","DOI":"10.1016\/j.chom.2012.08.002","type":"journal-article","article-title":"Clathrin Potentiates Vaccinia-Induced Actin Polymerization to Facilitate Viral Spread","volume":"12","author":"Humphries","year":"2012","unstructured":"Humphries, A. C. et al. Clathrin Potentiates Vaccinia-Induced Actin Polymerization to Facilitate Viral Spread. Cell Host Microbe. 12, 346-359 (2012).","journal-title":"Cell Host Microbe"},{"key":"ref36","doi-asserted-by":"crossref","first-page":"132","DOI":"10.1016\/j.jviromet.2012.07.003","type":"journal-article","article-title":"Sub-viral imaging of vaccinia virus using super-resolution microscopy","volume":"186","author":"Horsington","year":"2012","unstructured":"Horsington, J., Turnbull, L., Whitchurch, C. B. & Newsome, T. P. Sub-viral imaging of vaccinia virus using super-resolution microscopy. J. Virol. Methods. 186, 132-136 (2012).","journal-title":"J. Virol. Methods"},{"key":"ref37","doi-asserted-by":"crossref","DOI":"10.1371\/journal.ppat.1003239","type":"journal-article","article-title":"A36-dependent Actin Filament Nucleation Promotes Release of Vaccinia Virus","volume":"9","author":"Horsington","year":"2013","unstructured":"Horsington, J. et al. A36-dependent Actin Filament Nucleation Promotes Release of Vaccinia Virus. PLoS Pathog. 9, e1003239 (2013).","journal-title":"PLoS Pathog"}],"container-title":["Journal of Visualized Experiments"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/www.jove.com\/video\/51151\/methodology-for-efficient-generation-fluorescently-tagged-vaccinia","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T18:25:16Z","timestamp":1792693516000},"score":1,"resource":{"primary":{"URL":"https:\/\/app.jove.com\/t\/51151"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2014,1,17]]},"references-count":37,"journal-issue":{"issue":"83","published-online":{"date-parts":[[2014]]}},"URL":"https:\/\/doi.org\/10.3791\/51151","relation":{},"ISSN":["1940-087X"],"issn-type":[{"value":"1940-087X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2014,1,17]]}}}