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Here we describe a two-component genome yellow fever virus (YFV) replication system in which each of the genomes encodes complete sets of nonstructural proteins that form the replication complex but expresses either only capsid or prM\/E instead of the entire structural polyprotein. Upon delivery to the same cell, these genomes produce together all of the viral structural proteins, and cells release a combination of virions with both types of genomes packaged into separate particles. In tissue culture, this modified YFV can be further passaged at an escalating scale by using a high multiplicity of infection (MOI). However, at a low MOI, only one of the genomes is delivered into the cells, and infection cannot spread. The replicating prM\/E-encoding genome produces extracellular E protein in the form of secreted subviral particles that are known to be an effective immunogen. The presented strategy of developing viruses defective in replication might be applied to other flaviviruses, and these two-component genome viruses can be useful for diagnostic or vaccine applications, including the delivery and expression of heterologous genes. In addition, the achieved separation of the capsid-coding sequence and the cyclization signal in the YFV genome provides a new means for studying the mechanism of the flavivirus packaging process.<\/jats:p>","DOI":"10.1128\/jvi.01112-07","type":"journal-article","created":{"date-parts":[[2007,8,23]],"date-time":"2007-08-23T00:43:54Z","timestamp":1187829834000},"page":"11737-11748","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":49,"title":["Production of Pseudoinfectious Yellow Fever Virus with a Two-Component Genome"],"prefix":"10.1128","volume":"81","author":[{"given":"Alexandr V.","family":"Shustov","sequence":"first","affiliation":[{"name":"Department of Microbiology and Immunology"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter W.","family":"Mason","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology"},{"name":"Department of Pathology"},{"name":"Sealy Center for Vaccine Development, University of Texas Medical Branch, 301 University Blvd., Galveston, Texas 77555"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ilya","family":"Frolov","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"crossref","first-page":"6756","DOI":"10.4049\/jimmunol.163.12.6756","volume":"163","year":"1999","unstructured":"Aberle, J. 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