Characterisation of a new infectious full-length cDNA clone of BVDV genotype 2 and generation of virus mutants
Based on their genomic sequences, two genotypes of Bovine viral diarrhea virus (BVDV) can be differentiated, BVDV type 1 (BVDV-1) and BVDV type 2 (BVDV-2). The complete genomic sequence of the highly virulent BVDV-2 strain 890 was cloned as cDNA to establish the infectious cDNA clone p890FL. In vitr...
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| creator | Mischkale, Katrin Reimann, Ilona Zemke, J. König, P. Beer, Martin |
| description | Based on their genomic sequences, two genotypes of Bovine viral diarrhea virus (BVDV) can be differentiated, BVDV type 1 (BVDV-1) and BVDV type 2 (BVDV-2). The complete genomic sequence of the highly virulent BVDV-2 strain 890 was cloned as cDNA to establish the infectious cDNA clone p890FL. In vitro-synthesised full-length RNA of p890FL was transfected into bovine cells and infectious virus could be recovered (v890FL). In vitro, recombinant v890FL showed similar growth characteristics as wild type virus v890WT. However, infection experiments in calves revealed an attenuation of recombinant v890FL in comparison to the parental isolate. Both leukocytopenia and fever were less pronounced in v890FL-infected calves. Nevertheless, viremia and virus shedding were comparable between recombinant and parental BVDV 890. Furthermore, mutants with partial deletions of the genomic region encoding for the autoprotease Npro (p890ΔNpro) or the capsid protein (p890ΔC) were constructed and characterised. In order to generate pseudovirions, replicon p890ΔC was efficiently trans-complemented on a helper cell line. In summary, the newly developed construct p890FL represents the first infectious full-length cDNA clone for the BVDV-2 strain 890 and offers a useful tool for further studies on the pathogenesis of BVDV-2 and the development of novel recombinant BVDV-2 specific vaccine candidates. |
| doi_str_mv | 10.1016/j.vetmic.2009.09.036 |
| format | Article |
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The complete genomic sequence of the highly virulent BVDV-2 strain 890 was cloned as cDNA to establish the infectious cDNA clone p890FL. In vitro-synthesised full-length RNA of p890FL was transfected into bovine cells and infectious virus could be recovered (v890FL). In vitro, recombinant v890FL showed similar growth characteristics as wild type virus v890WT. However, infection experiments in calves revealed an attenuation of recombinant v890FL in comparison to the parental isolate. Both leukocytopenia and fever were less pronounced in v890FL-infected calves. Nevertheless, viremia and virus shedding were comparable between recombinant and parental BVDV 890. Furthermore, mutants with partial deletions of the genomic region encoding for the autoprotease Npro (p890ΔNpro) or the capsid protein (p890ΔC) were constructed and characterised. In order to generate pseudovirions, replicon p890ΔC was efficiently trans-complemented on a helper cell line. In summary, the newly developed construct p890FL represents the first infectious full-length cDNA clone for the BVDV-2 strain 890 and offers a useful tool for further studies on the pathogenesis of BVDV-2 and the development of novel recombinant BVDV-2 specific vaccine candidates.</description><identifier>ISSN: 0378-1135</identifier><identifier>EISSN: 1873-2542</identifier><identifier>DOI: 10.1016/j.vetmic.2009.09.036</identifier><identifier>PMID: 19875251</identifier><identifier>CODEN: VMICDQ</identifier><language>eng</language><publisher>Amsterdam: Elsevier B.V</publisher><subject>Animals ; Biological and medical sciences ; bovine viral diarrhea ; Bovine viral diarrhea virus ; Bovine viral diarrhea virus 1 ; Bovine viral diarrhea virus 2 ; Bovine viral diarrhea virus type 2 ; calves ; Cattle ; Cattle Diseases - pathology ; Cattle Diseases - virology ; Cell Line ; Cloning, Molecular ; complementary DNA ; Diarrhea Virus 2, Bovine Viral - genetics ; Diarrhea Virus 2, Bovine Viral - pathogenicity ; disease severity ; DNA, Complementary - genetics ; Fundamental and applied biological sciences. Psychology ; Genotype ; Hemorrhagic Syndrome, Bovine - pathology ; Hemorrhagic Syndrome, Bovine - veterinary ; Hemorrhagic Syndrome, Bovine - virology ; Infectious pestivirus clone ; Leukocyte Count ; leukopenia ; messenger RNA ; microbial genetics ; Microbiology ; Miscellaneous ; molecular sequence data ; mutants ; nucleotide sequences ; pathogen shedding ; pathogenesis ; Pestivirus ; Random Allocation ; Sequence Deletion - genetics ; signs and symptoms (animals and humans) ; strain differences ; strains ; viremia ; Virology ; virulence</subject><ispartof>Veterinary microbiology, 2010-04, Vol.142 (1-2), p.3-12</ispartof><rights>2009 Elsevier B.V.</rights><rights>2015 INIST-CNRS</rights><rights>Copyright 2009 Elsevier B.V. All rights reserved.</rights><rights>Distributed under a Creative Commons Attribution 4.0 International License</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c527t-ae7177c587ec171f2cac4b306fc0dd66efd5bda3dc0ff39bbd88e24aced7d2e83</citedby><cites>FETCH-LOGICAL-c527t-ae7177c587ec171f2cac4b306fc0dd66efd5bda3dc0ff39bbd88e24aced7d2e83</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0378113509004532$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>230,309,310,314,776,780,785,786,881,3536,23910,23911,25119,27903,27904,65309</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=22728737$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/19875251$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://hal.science/hal-00578395$$DView record in HAL$$Hfree_for_read</backlink></links><search><creatorcontrib>Mischkale, Katrin</creatorcontrib><creatorcontrib>Reimann, Ilona</creatorcontrib><creatorcontrib>Zemke, J.</creatorcontrib><creatorcontrib>König, P.</creatorcontrib><creatorcontrib>Beer, Martin</creatorcontrib><title>Characterisation of a new infectious full-length cDNA clone of BVDV genotype 2 and generation of virus mutants</title><title>Veterinary microbiology</title><addtitle>Vet Microbiol</addtitle><description>Based on their genomic sequences, two genotypes of Bovine viral diarrhea virus (BVDV) can be differentiated, BVDV type 1 (BVDV-1) and BVDV type 2 (BVDV-2). The complete genomic sequence of the highly virulent BVDV-2 strain 890 was cloned as cDNA to establish the infectious cDNA clone p890FL. In vitro-synthesised full-length RNA of p890FL was transfected into bovine cells and infectious virus could be recovered (v890FL). In vitro, recombinant v890FL showed similar growth characteristics as wild type virus v890WT. However, infection experiments in calves revealed an attenuation of recombinant v890FL in comparison to the parental isolate. Both leukocytopenia and fever were less pronounced in v890FL-infected calves. Nevertheless, viremia and virus shedding were comparable between recombinant and parental BVDV 890. Furthermore, mutants with partial deletions of the genomic region encoding for the autoprotease Npro (p890ΔNpro) or the capsid protein (p890ΔC) were constructed and characterised. In order to generate pseudovirions, replicon p890ΔC was efficiently trans-complemented on a helper cell line. In summary, the newly developed construct p890FL represents the first infectious full-length cDNA clone for the BVDV-2 strain 890 and offers a useful tool for further studies on the pathogenesis of BVDV-2 and the development of novel recombinant BVDV-2 specific vaccine candidates.</description><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>bovine viral diarrhea</subject><subject>Bovine viral diarrhea virus</subject><subject>Bovine viral diarrhea virus 1</subject><subject>Bovine viral diarrhea virus 2</subject><subject>Bovine viral diarrhea virus type 2</subject><subject>calves</subject><subject>Cattle</subject><subject>Cattle Diseases - pathology</subject><subject>Cattle Diseases - virology</subject><subject>Cell Line</subject><subject>Cloning, Molecular</subject><subject>complementary DNA</subject><subject>Diarrhea Virus 2, Bovine Viral - genetics</subject><subject>Diarrhea Virus 2, Bovine Viral - pathogenicity</subject><subject>disease severity</subject><subject>DNA, Complementary - genetics</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genotype</subject><subject>Hemorrhagic Syndrome, Bovine - pathology</subject><subject>Hemorrhagic Syndrome, Bovine - veterinary</subject><subject>Hemorrhagic Syndrome, Bovine - virology</subject><subject>Infectious pestivirus clone</subject><subject>Leukocyte Count</subject><subject>leukopenia</subject><subject>messenger RNA</subject><subject>microbial genetics</subject><subject>Microbiology</subject><subject>Miscellaneous</subject><subject>molecular sequence data</subject><subject>mutants</subject><subject>nucleotide sequences</subject><subject>pathogen shedding</subject><subject>pathogenesis</subject><subject>Pestivirus</subject><subject>Random Allocation</subject><subject>Sequence Deletion - genetics</subject><subject>signs and symptoms (animals and humans)</subject><subject>strain differences</subject><subject>strains</subject><subject>viremia</subject><subject>Virology</subject><subject>virulence</subject><issn>0378-1135</issn><issn>1873-2542</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkU1v0zAYgCMEYt3gHyDwBU0cUvyR2MkFqXTAkCo4wHa1HPt16ypxOjsp2r_HIVW5ISRLll8975efLHtF8JJgwt_vl0cYOqeXFON6OR3Gn2QLUgmW07KgT7MFZqLKCWHlRXYZ4x5jXNQcP88uSF2JkpZkkfn1TgWlBwguqsH1HvUWKeThF3Legk6hMSI7tm3egt8OO6Rvvq2QbnsPE_rx_uYebcH3w-MBEEXKm-kJ4Vzs6EKq0I2D8kN8kT2zqo3w8nRfZXefP_1c3-ab71--rlebXJdUDLkCQYTQZSVAE0Es1UoXDcPcamwM52BN2RjFjMbWsrppTFUBLZQGIwyFil1l7-a6O9XKQ3CdCo-yV07erjZyimFciorV5ZEk9npmD6F_GCEOsnNRQ9sqD2l5KQpOSk6q_yAZ40VBCpHIYiZ16GMMYM9DECwnfXIvZ31y0ienw3hKe31qMDYdmL9JJ18JeHsCVNSqtUF57eKZo1TQ5H_q_2bmrOql2ia58u4HxYThtEfN_-zyYSYgaTg6CDJqBz79oAtJuzS9-_esvwFUacQ4</recordid><startdate>20100421</startdate><enddate>20100421</enddate><creator>Mischkale, Katrin</creator><creator>Reimann, Ilona</creator><creator>Zemke, J.</creator><creator>König, P.</creator><creator>Beer, Martin</creator><general>Elsevier B.V</general><general>Amsterdam; New York: Elsevier</general><general>Elsevier</general><scope>FBQ</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>7TM</scope><scope>7U9</scope><scope>H94</scope><scope>1XC</scope><scope>VOOES</scope></search><sort><creationdate>20100421</creationdate><title>Characterisation of a new infectious full-length cDNA clone of BVDV genotype 2 and generation of virus mutants</title><author>Mischkale, Katrin ; Reimann, Ilona ; Zemke, J. ; König, P. ; Beer, Martin</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c527t-ae7177c587ec171f2cac4b306fc0dd66efd5bda3dc0ff39bbd88e24aced7d2e83</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>bovine viral diarrhea</topic><topic>Bovine viral diarrhea virus</topic><topic>Bovine viral diarrhea virus 1</topic><topic>Bovine viral diarrhea virus 2</topic><topic>Bovine viral diarrhea virus type 2</topic><topic>calves</topic><topic>Cattle</topic><topic>Cattle Diseases - pathology</topic><topic>Cattle Diseases - virology</topic><topic>Cell Line</topic><topic>Cloning, Molecular</topic><topic>complementary DNA</topic><topic>Diarrhea Virus 2, Bovine Viral - genetics</topic><topic>Diarrhea Virus 2, Bovine Viral - pathogenicity</topic><topic>disease severity</topic><topic>DNA, Complementary - genetics</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Genotype</topic><topic>Hemorrhagic Syndrome, Bovine - pathology</topic><topic>Hemorrhagic Syndrome, Bovine - veterinary</topic><topic>Hemorrhagic Syndrome, Bovine - virology</topic><topic>Infectious pestivirus clone</topic><topic>Leukocyte Count</topic><topic>leukopenia</topic><topic>messenger RNA</topic><topic>microbial genetics</topic><topic>Microbiology</topic><topic>Miscellaneous</topic><topic>molecular sequence data</topic><topic>mutants</topic><topic>nucleotide sequences</topic><topic>pathogen shedding</topic><topic>pathogenesis</topic><topic>Pestivirus</topic><topic>Random Allocation</topic><topic>Sequence Deletion - genetics</topic><topic>signs and symptoms (animals and humans)</topic><topic>strain differences</topic><topic>strains</topic><topic>viremia</topic><topic>Virology</topic><topic>virulence</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Mischkale, Katrin</creatorcontrib><creatorcontrib>Reimann, Ilona</creatorcontrib><creatorcontrib>Zemke, J.</creatorcontrib><creatorcontrib>König, P.</creatorcontrib><creatorcontrib>Beer, Martin</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Hyper Article en Ligne (HAL)</collection><collection>Hyper Article en Ligne (HAL) (Open Access)</collection><jtitle>Veterinary microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Mischkale, Katrin</au><au>Reimann, Ilona</au><au>Zemke, J.</au><au>König, P.</au><au>Beer, Martin</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Characterisation of a new infectious full-length cDNA clone of BVDV genotype 2 and generation of virus mutants</atitle><jtitle>Veterinary microbiology</jtitle><addtitle>Vet Microbiol</addtitle><date>2010-04-21</date><risdate>2010</risdate><volume>142</volume><issue>1-2</issue><spage>3</spage><epage>12</epage><pages>3-12</pages><issn>0378-1135</issn><eissn>1873-2542</eissn><coden>VMICDQ</coden><abstract>Based on their genomic sequences, two genotypes of Bovine viral diarrhea virus (BVDV) can be differentiated, BVDV type 1 (BVDV-1) and BVDV type 2 (BVDV-2). The complete genomic sequence of the highly virulent BVDV-2 strain 890 was cloned as cDNA to establish the infectious cDNA clone p890FL. In vitro-synthesised full-length RNA of p890FL was transfected into bovine cells and infectious virus could be recovered (v890FL). In vitro, recombinant v890FL showed similar growth characteristics as wild type virus v890WT. However, infection experiments in calves revealed an attenuation of recombinant v890FL in comparison to the parental isolate. Both leukocytopenia and fever were less pronounced in v890FL-infected calves. Nevertheless, viremia and virus shedding were comparable between recombinant and parental BVDV 890. Furthermore, mutants with partial deletions of the genomic region encoding for the autoprotease Npro (p890ΔNpro) or the capsid protein (p890ΔC) were constructed and characterised. In order to generate pseudovirions, replicon p890ΔC was efficiently trans-complemented on a helper cell line. In summary, the newly developed construct p890FL represents the first infectious full-length cDNA clone for the BVDV-2 strain 890 and offers a useful tool for further studies on the pathogenesis of BVDV-2 and the development of novel recombinant BVDV-2 specific vaccine candidates.</abstract><cop>Amsterdam</cop><pub>Elsevier B.V</pub><pmid>19875251</pmid><doi>10.1016/j.vetmic.2009.09.036</doi><tpages>10</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Animals Biological and medical sciences bovine viral diarrhea Bovine viral diarrhea virus Bovine viral diarrhea virus 1 Bovine viral diarrhea virus 2 Bovine viral diarrhea virus type 2 calves Cattle Cattle Diseases - pathology Cattle Diseases - virology Cell Line Cloning, Molecular complementary DNA Diarrhea Virus 2, Bovine Viral - genetics Diarrhea Virus 2, Bovine Viral - pathogenicity disease severity DNA, Complementary - genetics Fundamental and applied biological sciences. Psychology Genotype Hemorrhagic Syndrome, Bovine - pathology Hemorrhagic Syndrome, Bovine - veterinary Hemorrhagic Syndrome, Bovine - virology Infectious pestivirus clone Leukocyte Count leukopenia messenger RNA microbial genetics Microbiology Miscellaneous molecular sequence data mutants nucleotide sequences pathogen shedding pathogenesis Pestivirus Random Allocation Sequence Deletion - genetics signs and symptoms (animals and humans) strain differences strains viremia Virology virulence |
| title | Characterisation of a new infectious full-length cDNA clone of BVDV genotype 2 and generation of virus mutants |
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