Abstract:
:The complete nucleotide sequence of the genome of classical swine fever virus (CSFV) strain Alfort/187 was determined from three cDNA libraries constructed by cloning of DNA fragments obtained from independent sets of reverse transcription and PCR. The cDNA fragments were then assembled and inserted downstream of a T7 promoter in a P15A-derived plasmid vector to obtain the full-length cDNA clone pA187-1. The first nucleotide of the CSFV genome was positioned at the transcription start site of the T7 promoter. Cleavage at an SrfI restriction site introduced at the exact 3' end of the cloned viral cDNA allowed the in vitro synthesis of full-length viral RNA by runoff transcription. This RNA proved to be infectious after transfection into porcine kidney cells. Infectivity was not increased after capping of the synthetic RNA. Virus recovered from transfected cells was titrated in porcine kidney cells by endpoint dilution using indirect immunofluorescence and a CSFV-specific monoclonal antibody. RNA transcripts generated from plasmid DNA isolated from bacteria which had been cultured and cloned 10 times remained infectious, indicating that the full-length clone is stable in bacterial cells. A silent point mutation introduced at position 11842 of the genome was retained in the recombinant virus recovered from transfected cells. An infectious chimeric construct was obtained by replacing a 696-bp fragment in pA187-1 with the corresponding cDNA fragment from the CSFV strain CAP. The stably cloned full-length CSFV cDNA allows site-specific mutagenesis of the viral genome and thus will be useful for detailed molecular characterization of the virus as well as for studies of viral pathogenesis.
journal_name
J Viroljournal_title
Journal of virologyauthors
Ruggli N,Tratschin JD,Mittelholzer C,Hofmann MAdoi
10.1128/JVI.70.6.3478-3487.1996subject
Has Abstractpub_date
1996-06-01 00:00:00pages
3478-87issue
6eissn
0022-538Xissn
1098-5514journal_volume
70pub_type
杂志文章abstract::Based on sequence divergence in the transformation-relevant region, herpesvirus saimiri strains are classified into three subgroups. Only members of subgroup C transform human T lymphocytes to continuous interleukin-2-dependent growth in culture. In this study, human cord blood T cells were immortalized by using diffe...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.71.3.2252-2263.1997
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pub_type: 杂志文章
doi:10.1128/JVI.67.3.1357-1364.1993
更新日期:1993-03-01 00:00:00
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doi:10.1128/JVI.72.1.857-861.1998
更新日期:1998-01-01 00:00:00
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pub_type: 杂志文章
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更新日期:1994-12-01 00:00:00
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pub_type: 杂志文章
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更新日期:2006-05-01 00:00:00
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pub_type: 杂志文章
doi:10.1128/JVI.24.3.850-864.1977
更新日期:1977-12-01 00:00:00
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pub_type: 杂志文章
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更新日期:2014-06-01 00:00:00
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pub_type: 杂志文章
doi:10.1128/JVI.02726-06
更新日期:2007-06-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00640-07
更新日期:2007-10-01 00:00:00
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pub_type: 杂志文章,收录出版
doi:10.1128/JVI.02604-09
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pub_type: 杂志文章
doi:10.1128/JVI.17.3.841-853.1976
更新日期:1976-03-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00255-16
更新日期:2016-04-29 00:00:00
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pub_type: 杂志文章
doi:10.1128/JVI.01024-16
更新日期:2016-11-14 00:00:00
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更新日期:2011-11-01 00:00:00
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pub_type: 杂志文章
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更新日期:2021-01-27 00:00:00
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pub_type: 杂志文章
doi:10.1128/JVI.63.3.1188-1193.1989
更新日期:1989-03-01 00:00:00
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journal_title:Journal of virology
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更新日期:2004-09-01 00:00:00
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更新日期:2005-02-01 00:00:00
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更新日期:2003-09-01 00:00:00
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pub_type: 杂志文章
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更新日期:1986-06-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.16.4.1027-1038.1975
更新日期:1975-10-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.30.1.116-123.1979
更新日期:1979-04-01 00:00:00
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pub_type: 杂志文章
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更新日期:1998-03-01 00:00:00