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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Compatibility among Polymerase Subunit Proteins Is a Restricting Factor in Reassortment between Equine H7N7 and Human H3N2 Influenza Viruses<xref ref-type="fn" rid="fn1">▿</xref> </title><author><name sortKey="Li, Chengjun" sort="Li, Chengjun" uniqKey="Li C" first="Chengjun" last="Li">Chengjun Li</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Hatta, Masato" sort="Hatta, Masato" uniqKey="Hatta M" first="Masato" last="Hatta">Masato Hatta</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Watanabe, Shinji" sort="Watanabe, Shinji" uniqKey="Watanabe S" first="Shinji" last="Watanabe">Shinji Watanabe</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Neumann, Gabriele" sort="Neumann, Gabriele" uniqKey="Neumann G" first="Gabriele" last="Neumann">Gabriele Neumann</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Kawaoka, Yoshihiro" sort="Kawaoka, Yoshihiro" uniqKey="Kawaoka Y" first="Yoshihiro" last="Kawaoka">Yoshihiro Kawaoka</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">18815312</idno><idno type="pmc">2583690</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2583690</idno><idno type="RBID">PMC:2583690</idno><idno type="doi">10.1128/JVI.01445-08</idno><date when="2008">2008</date><idno type="wicri:Area/Pmc/Corpus">000365</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000365</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Compatibility among Polymerase Subunit Proteins Is a Restricting Factor in Reassortment between Equine H7N7 and Human H3N2 Influenza Viruses<xref ref-type="fn" rid="fn1">▿</xref> </title><author><name sortKey="Li, Chengjun" sort="Li, Chengjun" uniqKey="Li C" first="Chengjun" last="Li">Chengjun Li</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Hatta, Masato" sort="Hatta, Masato" uniqKey="Hatta M" first="Masato" last="Hatta">Masato Hatta</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Watanabe, Shinji" sort="Watanabe, Shinji" uniqKey="Watanabe S" first="Shinji" last="Watanabe">Shinji Watanabe</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Neumann, Gabriele" sort="Neumann, Gabriele" uniqKey="Neumann G" first="Gabriele" last="Neumann">Gabriele Neumann</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Kawaoka, Yoshihiro" sort="Kawaoka, Yoshihiro" uniqKey="Kawaoka Y" first="Yoshihiro" last="Kawaoka">Yoshihiro Kawaoka</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></analytic><series><title level="j">Journal of Virology</title><idno type="ISSN">0022-538X</idno><idno type="eISSN">1098-5514</idno><imprint><date when="2008">2008</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Reassortment is an important driving force for influenza virus evolution, and a better understanding of the factors that affect this process could improve our ability to respond to future influenza pandemics and epidemics. To identify factors that restrict the generation of reassortant viruses, we cotransfected human embryonic kidney cells with plasmids for the synthesis of viral RNAs of both A/equine/Prague/1/56 (Prague; H7N7) and A/Yokohama/2017/03 (Yokohama; H3N2) viruses together with the supporting protein expression plasmids. Of the possible 256 genotypes, we identified 29 genotypes in 120 randomly plaque-picked reassortants examined. Analyses of these reassortants suggested that the formation of functional ribonucleoprotein (RNP) complexes was a restricting factor, a finding that correlated with the activities of RNP complexes composed of different combinations of the proteins from the two viruses, as measured in a minigenome assay. For at least one nonfunctional RNP complex (i.e., Prague PB2, Prague PB1, Yokohama PA, and Prague NP), the lack of activity was due to the inability of the three polymerase subunit proteins to form a heterotrimer. Adaptation of viruses possessing a gene encoding a chimera of the PA proteins of the two viruses and the remaining genes from Prague virus resulted in compensatory mutations in the PB2 and/or PA protein. These results indicate substantial incompatibility among the gene products of the two test viruses, a critical role for the RNP complex in the generation of reassortant viruses, and a functional interaction of PB2 and PA.</p></div></front></TEI><pmc article-type="research-article"><pmc-comment>The publisher of this article does not allow downloading of the full text in XML form.</pmc-comment>
<front><journal-meta><journal-id journal-id-type="nlm-ta">J Virol</journal-id><journal-id journal-id-type="publisher-id">jvi</journal-id><journal-title>Journal of Virology</journal-title><issn pub-type="ppub">0022-538X</issn><issn pub-type="epub">1098-5514</issn><publisher><publisher-name>American Society for Microbiology (ASM)</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">18815312</article-id><article-id pub-id-type="pmc">2583690</article-id><article-id pub-id-type="publisher-id">1445-08</article-id><article-id pub-id-type="doi">10.1128/JVI.01445-08</article-id><article-categories><subj-group subj-group-type="heading"><subject>Genetic Diversity and Evolution</subject></subj-group></article-categories><title-group><article-title>Compatibility among Polymerase Subunit Proteins Is a Restricting Factor in Reassortment between Equine H7N7 and Human H3N2 Influenza Viruses<xref ref-type="fn" rid="fn1">▿</xref> </article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Li</surname><given-names>Chengjun</given-names></name><xref ref-type="aff" rid="aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Hatta</surname><given-names>Masato</given-names></name><xref ref-type="aff" rid="aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Watanabe</surname><given-names>Shinji</given-names></name><xref ref-type="aff" rid="aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Neumann</surname><given-names>Gabriele</given-names></name><xref ref-type="aff" rid="aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Kawaoka</surname><given-names>Yoshihiro</given-names></name><xref ref-type="aff" rid="aff1">1</xref><xref ref-type="aff" rid="aff1">2</xref><xref ref-type="aff" rid="aff1">3</xref><xref ref-type="corresp" rid="cor1">*</xref></contrib></contrib-group><aff id="aff1">Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, 2015 Linden Drive, Madison, Wisconsin 53706,<label>1</label> Division of Virology, Department of Microbiology and Immunology,<label>2</label> International Research Center for Infectious Diseases, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, Japan<label>3</label></aff><author-notes><fn id="cor1"><label>*</label><p>Corresponding author. Mailing address: Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, 2015 Linden Drive, Madison, WI 53706. Phone: (608) 265-4925. Fax: (608) 265-5622. E-mail: <email>kawaokay@svm.vetmed.wisc.edu</email></p></fn></author-notes><pub-date pub-type="ppub"><month>12</month><year>2008</year></pub-date><pub-date pub-type="epub"><day>24</day><month>9</month><year>2008</year></pub-date><volume>82</volume><issue>23</issue><fpage>11880</fpage><lpage>11888</lpage><history><date date-type="received"><day>10</day><month>7</month><year>2008</year></date><date date-type="accepted"><day>11</day><month>9</month><year>2008</year></date></history><permissions><copyright-statement>Copyright © 2008, American Society for Microbiology</copyright-statement></permissions><self-uri xlink:title="pdf" xlink:href="zjv02308011880.pdf"></self-uri><abstract><p>Reassortment is an important driving force for influenza virus evolution, and a better understanding of the factors that affect this process could improve our ability to respond to future influenza pandemics and epidemics. To identify factors that restrict the generation of reassortant viruses, we cotransfected human embryonic kidney cells with plasmids for the synthesis of viral RNAs of both A/equine/Prague/1/56 (Prague; H7N7) and A/Yokohama/2017/03 (Yokohama; H3N2) viruses together with the supporting protein expression plasmids. Of the possible 256 genotypes, we identified 29 genotypes in 120 randomly plaque-picked reassortants examined. Analyses of these reassortants suggested that the formation of functional ribonucleoprotein (RNP) complexes was a restricting factor, a finding that correlated with the activities of RNP complexes composed of different combinations of the proteins from the two viruses, as measured in a minigenome assay. For at least one nonfunctional RNP complex (i.e., Prague PB2, Prague PB1, Yokohama PA, and Prague NP), the lack of activity was due to the inability of the three polymerase subunit proteins to form a heterotrimer. Adaptation of viruses possessing a gene encoding a chimera of the PA proteins of the two viruses and the remaining genes from Prague virus resulted in compensatory mutations in the PB2 and/or PA protein. These results indicate substantial incompatibility among the gene products of the two test viruses, a critical role for the RNP complex in the generation of reassortant viruses, and a functional interaction of PB2 and PA.</p></abstract></article-meta></front><floats-wrap><fig position="float" id="f1"><label>FIG. 1.</label><caption><p>Activities of 16 RNP combinations measured in a minigenome assay. Four protein expression plasmids (PB2, PB1, PA, and NP [0.5 μg of each]) for the 16 RNP combinations were transfected into 293 cells together with pPolWSNNA F-Luc, which encodes a vRNA possessing a reporter firefly luciferase gene. At 48 h posttransfection, a dual-luciferase assay was performed in which the relative firefly luciferase activity was normalized to the internal control, <italic>Renilla</italic> luciferase activity (see Materials and Methods). The values shown are means ± standard deviations for three independent experiments and are standardized to the activity of P<sub>PB2</sub>P<sub>PB1</sub>P<sub>PA</sub>P<sub>NP</sub> (100%). P, Prague. Y, Yokohama.</p></caption><graphic xlink:href="zjv0230812910001"></graphic></fig><fig position="float" id="f2"><label>FIG. 2.</label><caption><p>Interactions among RNA polymerase subunits. (A) 293T cells were cotransfected with 2 μg each of protein expression plasmids, as indicated (+). At 48 h posttransfection, cell lysates were incubated with a goat polyclonal anti-PB2 antibody (Santa Cruz) (lanes 1 to 4). The agarose bead-bound proteins were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotted with mouse monoclonal anti-PB2, anti-PB1, or anti-FLAG antibodies. The cell lysates were also directly Western blotted with mouse monoclonal anti-PB2, anti-PB1, and anti-FLAG antibodies, as shown in lanes 5 to 8. (B) 293T cells were transfected with 2 μg of protein expression plasmids encoding Prague PB2, Prague PB1, Yokohama PB1, Prague PA, and Yokohama PA. At 48 h posttransfection, cell lysates were prepared and Western blotted with mouse monoclonal antibodies (anti-PB2 [lanes 1 and 2], anti-PB1 [lanes 3 to 5], and anti-FLAG [lanes 6 to 8]).</p></caption><graphic xlink:href="zjv0230812910002"></graphic></fig><fig position="float" id="f3"><label>FIG. 3.</label><caption><p>Schematic diagram of Prague/Yokohama PA chimeras and their effects on RNP activities in a minigenome assay. The 12 PA chimeras were constructed as described in Materials and Methods. The noncoding regions of Prague and Yokohama PA proteins are represented by light blue and pink bars, respectively. The coding regions of Prague and Yokohama PA proteins are shown by blue and red bars, respectively. All constructs are shown in the negative-sense orientation. The RNP combination P<sub>PB2</sub>P<sub>PB1</sub>P<sub>(Y)PA</sub>P<sub>NP</sub> [Prague PB2 and PB1, chimeric PA, and Prague NP; P<sub>(Y)PA</sub>, chimera of Prague and Yokohama PA] was tested in the minigenome assay described in Materials and Methods. The relative firefly luciferase activity was normalized to the <italic>Renilla</italic> luciferase activity, which served as the internal control. The activity of the RNP combination P<sub>PB2</sub>P<sub>PB1</sub>P<sub>PA</sub>P<sub>NP</sub> was standardized to 100%. The error bars indicate the standard deviations for three independent experiments. P, Prague. Y, Yokohama. P<sub>PB2</sub>P<sub>PB1</sub>P<sub>(Y1-100)PA</sub>P<sub>NP</sub>, RNP combination with Prague PB2, Prague PB1, Pr<sub>(Yok 1-100)</sub>PA, and Prague NP. Other RNP combinations were similarly named.</p></caption><graphic xlink:href="zjv0230812910003"></graphic></fig><fig position="float" id="f4"><label>FIG. 4.</label><caption><p>Activities of P<sub>PB2</sub>P<sub>PB1</sub>P<sub>(Y)PA</sub>P<sub>NP</sub> RNP combinations possessing mutations in PB2 or the chimeric PA. Four protein expression plasmids (0.5 μg each), including one for Prague PB2 (or mutant PB2), Prague PB1, PA chimera (or mutant PA chimera), and Prague NP, were transfected into 293 cells together with pPolWSNNA F-Luc. At 48 h posttransfection, luciferase assay was performed with a dual-luciferase kit. Firefly luciferase activity is shown as log values. 1-400, RNP combination with Prague PB2, Prague PB1, Pr<sub>(Yok 1-400)</sub>PA, and Prague NP; D253N, RNP combination with Prague PB2/D253N, Prague PB1, Pr<sub>(Yok 1-400)</sub>PA, and Prague NP. Other RNP combinations were similarly named. Mutations in the PB2 gene are shown in red; those in the PA gene are shown in blue. P, Prague.</p></caption><graphic xlink:href="zjv0230812910004"></graphic></fig><table-wrap position="float" id="t1"><label>TABLE 1.</label><caption><p>Genotypes of reassortant viruses between Prague (H7N7) and Yokohama (H3N2) viruses<xref ref-type="table-fn" rid="t1fn1"><italic>a</italic></xref></p></caption><table frame="hsides" rules="groups"><thead><tr><th colspan="1" rowspan="2" align="center" valign="middle">Genotype</th><th colspan="8" rowspan="1" align="center" valign="bottom">Gene origin<xref ref-type="table-fn" rid="t1fn2"><italic>b</italic></xref><hr></hr></th><th colspan="1" rowspan="2" align="center" valign="middle">Frequency (no. of viruses)</th></tr><tr><th colspan="1" rowspan="1" align="center" valign="bottom">PB2</th><th colspan="1" rowspan="1" align="center" valign="bottom">PB1</th><th colspan="1" rowspan="1" align="center" valign="bottom">PA</th><th colspan="1" rowspan="1" align="center" valign="bottom">HA</th><th colspan="1" rowspan="1" align="center" valign="bottom">NP</th><th colspan="1" rowspan="1" align="center" valign="bottom">NA</th><th colspan="1" rowspan="1" align="center" valign="bottom">M</th><th colspan="1" rowspan="1" align="center" valign="bottom">NS</th></tr></thead><tbody><tr><td colspan="1" rowspan="1" align="left" valign="top">A1</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="char" char="." valign="top">6</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A2</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="char" char="." valign="top">5</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A3</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="char" char="." valign="top">4</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A4</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="char" char="." valign="top">11</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A5</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A6</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="char" char="." valign="top">23</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A7</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="char" char="." valign="top">6</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A8</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="char" char="." valign="top">2</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A9</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="char" char="." valign="top">4</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A10</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="char" char="." valign="top">14</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A11</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A12</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A13</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="char" char="." valign="top">14</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A14</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A15</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="char" char="." valign="top">2</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A16</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A17</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A18</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A19</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A20</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="char" char="." valign="top">2</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A21</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A22</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A23</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="char" char="." valign="top">4</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A24</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="char" char="." valign="top">4</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A25</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A26</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A27</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A28</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">A29</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">Y</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="left" valign="top">P</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">No. of viruses possessing the respective Yokohama segment/total no. of viruses</td><td colspan="1" rowspan="1" align="left" valign="top">120/120</td><td colspan="1" rowspan="1" align="left" valign="top">112/120</td><td colspan="1" rowspan="1" align="left" valign="top">86/120</td><td colspan="1" rowspan="1" align="left" valign="top">118/120</td><td colspan="1" rowspan="1" align="left" valign="top">15/120</td><td colspan="1" rowspan="1" align="left" valign="top">76/120</td><td colspan="1" rowspan="1" align="left" valign="top">31/120</td><td colspan="1" rowspan="1" align="left" valign="top">83/120</td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr></tbody></table><table-wrap-foot><fn id="t1fn1"><label>a</label><p>293T cells were transfected with plasmids for the generation of vRNAs for both the Prague and Yokohama viruses together with the four protein expression plasmids of WSN. The cotransfection supernatants were plaque purified on MDCK cells. The genotypes of the plaque-purified viruses were determined by using strain-specific PCRs with identifying primers.</p></fn><fn id="t1fn2"><label>b</label><p>P, Prague; Y, Yokohama.</p></fn></table-wrap-foot></table-wrap><table-wrap position="float" id="t2"><label>TABLE 2.</label><caption><p>Gene combinations of RNP complexes in reassortant viruses between Prague (H7N7) and Yokohama (H3N2) viruses<xref ref-type="table-fn" rid="t2fn1"><italic>a</italic></xref></p></caption><table frame="hsides" rules="groups"><thead><tr><th colspan="1" rowspan="2" align="center" valign="middle">Combination no.</th><th colspan="4" rowspan="1" align="center" valign="bottom">RNP gene origin
<hr></hr></th><th colspan="1" rowspan="2" align="center" valign="middle">Frequency (no. of reassortant viruses)</th></tr><tr><th colspan="1" rowspan="1" align="center" valign="bottom">PB2</th><th colspan="1" rowspan="1" align="center" valign="bottom">PB1</th><th colspan="1" rowspan="1" align="center" valign="bottom">PA</th><th colspan="1" rowspan="1" align="center" valign="bottom">NP</th></tr></thead><tbody><tr><td colspan="1" rowspan="1" align="left" valign="top">1</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">7</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">2</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">−</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">3</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">−</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">4</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">−</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">5</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">25</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">6</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">−</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">7</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">1</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">8</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">−</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">9</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">−</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">10</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">−</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">11</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">73</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">12</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">1</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">13</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">−</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">14</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">−</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">15</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">Y</td><td colspan="1" rowspan="1" align="center" valign="top">13</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">16</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">P</td><td colspan="1" rowspan="1" align="center" valign="top">−</td></tr></tbody></table><table-wrap-foot><fn id="t2fn1"><label>a</label><p>Frequencies are based on the characterization of 120 plaque-purified viruses (see Table <xref ref-type="table" rid="t1">1</xref>). P, Prague; Y, Yokohama. −, the RNP gene combination was not present among the plaque-purified viruses.</p></fn></table-wrap-foot></table-wrap><table-wrap position="float" id="t3"><label>TABLE 3.</label><caption><p>Rescue of Prague viruses with PA chimeras<xref ref-type="table-fn" rid="t3fn1"><italic>a</italic></xref></p></caption><table frame="hsides" rules="groups"><thead><tr><th colspan="1" rowspan="2" align="center" valign="middle">Virus<xref ref-type="table-fn" rid="t3fn2"><italic>b</italic></xref></th><th colspan="2" rowspan="1" align="center" valign="bottom">Detection of virus in eggs
<hr></hr></th></tr><tr><th colspan="1" rowspan="1" align="center" valign="bottom">Passage 1</th><th colspan="1" rowspan="1" align="center" valign="bottom">Passage 2</th></tr></thead><tbody><tr><td colspan="1" rowspan="1" align="left" valign="top">Prague/Pr<sub>(Yok 1-100)</sub>PA</td><td colspan="1" rowspan="1" align="center" valign="top">+</td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Prague/Pr<sub>(Yok 1-200)</sub>PA</td><td colspan="1" rowspan="1" align="center" valign="top">+</td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Prague/Pr<sub>(Yok 1-300)</sub>PA</td><td colspan="1" rowspan="1" align="center" valign="top">+</td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Prague/Pr<sub>(Yok 1-400)</sub>PA</td><td colspan="1" rowspan="1" align="center" valign="top">+</td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Prague/Pr<sub>(Yok 1-500)</sub>PA</td><td colspan="1" rowspan="1" align="center" valign="top">−</td><td colspan="1" rowspan="1" align="center" valign="top">+</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Prague/Pr<sub>(Yok 1-600)</sub>PA</td><td colspan="1" rowspan="1" align="center" valign="top">−</td><td colspan="1" rowspan="1" align="center" valign="top">+</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Prague/Pr<sub>(Yok 101-716)</sub>PA</td><td colspan="1" rowspan="1" align="center" valign="top">−</td><td colspan="1" rowspan="1" align="center" valign="top">+</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Prague/Pr<sub>(Yok 201-716)</sub>PA</td><td colspan="1" rowspan="1" align="center" valign="top">−</td><td colspan="1" rowspan="1" align="center" valign="top">+</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Prague/Pr<sub>(Yok 301-716)</sub>PA</td><td colspan="1" rowspan="1" align="center" valign="top">−</td><td colspan="1" rowspan="1" align="center" valign="top">+</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Prague/Pr<sub>(Yok 401-716)</sub>PA</td><td colspan="1" rowspan="1" align="center" valign="top">+</td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Prague/Pr<sub>(Yok 501-716)</sub>PA</td><td colspan="1" rowspan="1" align="center" valign="top">+</td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Prague/Pr<sub>(Yok 601-716)</sub>PA</td><td colspan="1" rowspan="1" align="center" valign="top">+</td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Prague/Yok PA<xref ref-type="table-fn" rid="t3fn3"><italic>c</italic></xref></td><td colspan="1" rowspan="1" align="center" valign="top">−</td><td colspan="1" rowspan="1" align="center" valign="top">−</td></tr></tbody></table><table-wrap-foot><fn id="t3fn1"><label>a</label><p>Prague viruses possessing chimeric PA genes were generated as described in Materials and Methods. The transfection supernatant was inoculated into the allantoic cavities of 10-day-old chicken embryonated eggs. The presence of virus was confirmed by HA assay. A second passage in eggs was performed for those viruses that could not be detected after the first passage in eggs. +, virus was detected by HA assay; −, virus was not detected by HA assay.</p></fn><fn id="t3fn2"><label>b</label><p>The Prague viruses with PA chimeras were named according to the chimeric PAs they contained.</p></fn><fn id="t3fn3"><label>c</label><p>Single-gene reassortant Prague virus containing Yokohama PA.</p></fn></table-wrap-foot></table-wrap><table-wrap position="float" id="t4"><label>TABLE 4.</label><caption><p>Amino acid mutations in the RNP complexes of Prague viruses possessing chimeric PA genes<xref ref-type="table-fn" rid="t4fn1"><italic>a</italic></xref></p></caption><table frame="hsides" rules="groups"><thead><tr><th colspan="1" rowspan="2" align="center" valign="middle">Virus</th><th colspan="8" rowspan="1" align="center" valign="bottom">Presence of PB2 mutation
<hr></hr></th><th colspan="2" rowspan="1" align="center" valign="bottom">Presence of PA mutation
<hr></hr></th></tr><tr><th colspan="1" rowspan="1" align="center" valign="bottom">D253N</th><th colspan="1" rowspan="1" align="center" valign="bottom">D256G</th><th colspan="1" rowspan="1" align="center" valign="bottom">S279L</th><th colspan="1" rowspan="1" align="center" valign="bottom">T371I</th><th colspan="1" rowspan="1" align="center" valign="bottom">N425T</th><th colspan="1" rowspan="1" align="center" valign="bottom">H432R</th><th colspan="1" rowspan="1" align="center" valign="bottom">N540S</th><th colspan="1" rowspan="1" align="center" valign="bottom">N677T</th><th colspan="1" rowspan="1" align="center" valign="bottom">N444S</th><th colspan="1" rowspan="1" align="center" valign="bottom">A448V</th></tr></thead><tbody><tr><td colspan="1" rowspan="1" align="left" valign="top">Prague/Pr<sub>(Yok 1-100)</sub>PA</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Prague/Pr<sub>(Yok 1-200)</sub>PA</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Prague/Pr<sub>(Yok 1-300)</sub>PA</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Prague/Pr<sub>(Yok 1-400)</sub>PA</td><td colspan="1" rowspan="1" align="center" valign="top">+</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Prague/Pr<sub>(Yok 1-500)</sub>PA</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top">+</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top">+</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Prague/Pr<sub>(Yok 1-600)</sub>PA</td><td colspan="1" rowspan="1" align="center" valign="top">+</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top">+</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Prague/Pr<sub>(Yok 101-716)</sub>PA</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top">+</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top">+</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Prague/Pr<sub>(Yok 201-716)</sub>PA</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top">+</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Prague/Pr<sub>(Yok 301-716)</sub>PA</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top">+</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top">+</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Prague/Pr<sub>(Yok 401-716)</sub>PA</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top">+</td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Prague/Pr<sub>(Yok 501-716)</sub>PA</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Prague/Pr<sub>(Yok 601-716)</sub>PA</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr></tbody></table><table-wrap-foot><fn id="t4fn1"><label>a</label><p>The allantoic fluid of Prague viruses possessing chimeric PA genes was subjected to reverse transcription-PCR, and the PB2, PB1, PA, and NP genes were fully sequenced. The mutations found in the RNP complexes of the Prague chimeric PA viruses are shown. +, mutation present.</p></fn></table-wrap-foot></table-wrap></floats-wrap></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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