Abstract

Previously, cDNAs were constructed to encode various truncated versions of the genome RNA of respiratory syncytial virus (RSV) in which the genome termini and putative transcription signals were retained but the viral genes were deleted and replaced with a reporter gene such as that for bacterial chloramphenicol acetyl transferase (CAT). When introduced into RSV-infected cells, the prototype RSV-CAT minigenome was replicated, transcribed and packaged into virus-like particles. This represents the first type of system in which genome-like RNAs of a nonsegmented negative strand RNA virus could be introduced into the viral replication cycle and rendered biologically active. Here, this system was used to identify and characterize the cis-acting replication and transcription signals in the RSV genome and to study the mechanism of gene expression. It also was adapted for the analysis of protein function and for the generation of infectious RSV from a cDNA of the complete genome (accompanying reports).

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Intramural Research (Z01)
Project #
1Z01AI000368-13
Application #
5200456
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
13
Fiscal Year
1995
Total Cost
Indirect Cost

  Institution

Name
National Institute of Allergy and Infectious Diseases
Department
Type
DUNS #
City
State
Country
United States
Zip Code

  Publications

Melendi, Guillermina A; Zavala, Fidel; Buchholz, Ursula J et al. (2007) Mapping and characterization of the primary and anamnestic H-2(d)-restricted cytotoxic T-lymphocyte response in mice against human metapneumovirus. J Virol 81:11461-7
Buchholz, Ursula J; Nagashima, Kunio; Murphy, Brian R et al. (2006) Live vaccines for human metapneumovirus designed by reverse genetics. Expert Rev Vaccines 5:695-706
Biacchesi, Stephane; Pham, Quynh N; Skiadopoulos, Mario H et al. (2006) Modification of the trypsin-dependent cleavage activation site of the human metapneumovirus fusion protein to be trypsin independent does not increase replication or spread in rodents or nonhuman primates. J Virol 80:5798-806
Biacchesi, Stephane; Skiadopoulos, Mario H; Yang, Lijuan et al. (2005) Rapid human metapneumovirus microneutralization assay based on green fluorescent protein expression. J Virol Methods 128:192-7
Buchholz, Ursula J; Biacchesi, Stephane; Pham, Quynh N et al. (2005) Deletion of M2 gene open reading frames 1 and 2 of human metapneumovirus: effects on RNA synthesis, attenuation, and immunogenicity. J Virol 79:6588-97
Pham, Quynh N; Biacchesi, Stephane; Skiadopoulos, Mario H et al. (2005) Chimeric recombinant human metapneumoviruses with the nucleoprotein or phosphoprotein open reading frame replaced by that of avian metapneumovirus exhibit improved growth in vitro and attenuation in vivo. J Virol 79:15114-22
Biacchesi, Stephane; Skiadopoulos, Mario H; Yang, Lijuan et al. (2004) Recombinant human Metapneumovirus lacking the small hydrophobic SH and/or attachment G glycoprotein: deletion of G yields a promising vaccine candidate. J Virol 78:12877-87
Biacchesi, Stephane; Skiadopoulos, Mario H; Tran, Kim C et al. (2004) Recovery of human metapneumovirus from cDNA: optimization of growth in vitro and expression of additional genes. Virology 321:247-59
Buchholz, Ursula J; Bukreyev, Alexander; Yang, Lijuan et al. (2004) Contributions of the structural proteins of severe acute respiratory syndrome coronavirus to protective immunity. Proc Natl Acad Sci U S A 101:9804-9
Biacchesi, Stephane; Skiadopoulos, Mario H; Boivin, Guy et al. (2003) Genetic diversity between human metapneumovirus subgroups. Virology 315:1-9

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