Reverse genetics system for porcine enteric calicivirus, a prototype Sapovirus in the Caliciviridae

被引:57
作者
Chang, KO
Sosnovtsev, SS
Belliot, G
Wang, QH
Saif, LJ
Green, KY
机构
[1] NIAID, LID, Natl Inst Hlth, DHHS, Bethesda, MD 20892 USA
[2] Ohio State Univ, Dept Vet Prevent Med, Food Anim Hlth Res Program, Wooster, OH USA
关键词
D O I
10.1128/JVI.79.3.1409-1416.2005
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
A porcine enteric calicivirus (PEC), strain Cowden in the genus Sapovirus of the Caliciviridae family, can be propagated in a porcine kidney continuous cell line (LLC-PK) in the presence of bile acids in the cell culture medium. A full-length cDNA copy of the Cowden PEC genome was cloned into a plasmid vector directly downstream from the T7 RNA polymerase promoter, and capped RNA transcripts derived from this clone were infectious when transfected into LLC-PK cells. The recovery of PEC after transfection of RNA transcripts was dependent on the presence of bile acids, consistent with our recent identification of a bile acid-mediated signaling pathway required for PEC replication (Chang et al., Proc. Natl. Acad. Sci. USA 101:8733-87:88, 2004). Recovery of virus was verified by detection of PEC antigen in transfected cells by immunofluorescence and enzyme-linked immunosorbent assays, direct observation of recovered viral particles by electron microscopy, and partial sequence analysis of their genomes (first 1,070 nucleotides) to differentiate them from tissue culture-adapted parental virus. The recovered virus retained its ability to infect piglets when administered by the oral route and showed an attenuated phenotype similar to that of the tissue culture-adapted parental virus. This reverse genetics system for PEC provides a new tool to study the molecular basis of replication and pathogenesis for caliciviruses associated with diarrheal disease.
引用
收藏
页码:1409 / 1416
页数:8
相关论文
共 26 条
  • [1] Bile acids are essential for porcine enteric calicivirus replication in association with down-regulation of signal transducer and activator of transcription 1
    Chang, KO
    Sosnovtsev, SV
    Belliot, G
    Kim, Y
    Saif, LJ
    Green, KY
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2004, 101 (23) : 8733 - 8738
  • [2] Cell-culture propagation of porcine enteric calicivirus mediated by intestinal contents is dependent on the cyclic AMP signaling pathway
    Chang, KO
    Kim, Y
    Green, KY
    Saif, LJ
    [J]. VIROLOGY, 2002, 304 (02) : 302 - 310
  • [3] The molecular biology of caliciviruses
    Clarke, IN
    Lambden, PR
    [J]. JOURNAL OF GENERAL VIROLOGY, 1997, 78 : 291 - 301
  • [4] CUBITT D, 1994, VIRAL INFECT GASTROI, P549
  • [5] Molecular epidemiology of "Norwalk-like viruses" in outbreaks of gastroenteritis in the united states
    Fankhauser, RL
    Noel, JS
    Monroe, SS
    Ando, T
    Glass, RI
    [J]. JOURNAL OF INFECTIOUS DISEASES, 1998, 178 (06) : 1571 - 1578
  • [6] SERIAL PROPAGATION OF PORCINE ENTERIC CALICIVIRUS-LIKE VIRUS IN PRIMARY PORCINE KIDNEY-CELL CULTURES
    FLYNN, WT
    SAIF, LJ
    [J]. JOURNAL OF CLINICAL MICROBIOLOGY, 1988, 26 (02) : 206 - 212
  • [7] FLYNN WT, 1988, AM J VET RES, V49, P819
  • [8] Genetic and antigenic heterogeneity among feline calicivirus isolates from distinct disease manifestations
    Geissler, K
    Schneider, K
    Platzer, G
    Truyen, B
    Kaaden, OR
    Truyen, U
    [J]. VIRUS RESEARCH, 1997, 48 (02) : 193 - 206
  • [9] Green K., 2001, FIELDS VIROLOGY, V2, P841
  • [10] Taxonomy of the caliciviruses
    Green, KY
    Ando, T
    Balayan, MS
    Berke, T
    Clarke, IN
    Estes, MK
    Matson, DO
    Nakata, S
    Neill, JD
    Studdert, MJ
    Thiel, HJ
    [J]. JOURNAL OF INFECTIOUS DISEASES, 2000, 181 : S322 - S330