Linked Life Data

16569439

Source:http://linkedlifedata.com/resource/pubmed/id/16569439

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
2006-5-23
pubmed:abstractText
Recovery of recombinant, negative-strand, nonsegmented RNA viruses from a genomic cDNA clone requires a rescue system that promotes de novo assembly of a functional ribonucleoprotein (RNP) complex in the cell cytoplasm. This is accomplished typically by cotransfecting permissive cells with multiple plasmids that encode the positive-sense genomic RNA, the nucleocapsid protein (N or NP), and the two subunits of the viral RNA-dependent RNA polymerase (L and P). The transfected plasmids are transcribed in the cell cytoplasm by phage T7 RNA polymerase (T7 RNAP), which usually is supplied by infection with a recombinant vaccinia virus or through use of a stable cell line that expresses the polymerase. Although both methods of providing T7 RNAP are effective neither is ideal for viral vaccine development for a number of reasons. Therefore, it was necessary to modify existing technology to make it possible to routinely rescue a variety of recombinant viruses when T7 RNAP was provided by a cotransfected expression plasmid. Development of a broadly applicable procedure required optimization of the helper-virus-free methodology, which resulted in several modifications that improved rescue efficiency such as inclusion of plasmids encoding viral glycoproteins and matrix protein, heat shock treatment, and use of electroporation. The combined effect of these enhancements produced several important benefits including: (1) a helper-virus-free methodology capable of rescuing a diverse variety of paramyxoviruses and recombinant vesicular stomatitis virus (rVSV); (2) methodology that functioned effectively when using Vero cells, a suitable substrate for vaccine production; and (3) a method that enabled rescue of highly attenuated recombinant viruses, which had proven refractory to rescue using published procedures.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jul
pubmed:issn
0166-0934
pubmed:author
pubmed:issnType
Print
pubmed:volume
135
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
91-101
pubmed:dateRevised
2007-11-15
pubmed:meshHeading
pubmed-meshheading:16569439-Animals, pubmed-meshheading:16569439-Cercopithecus aethiops, pubmed-meshheading:16569439-DNA, Recombinant, pubmed-meshheading:16569439-DNA, Viral, pubmed-meshheading:16569439-DNA-Directed RNA Polymerases, pubmed-meshheading:16569439-Helper Viruses, pubmed-meshheading:16569439-Mutation, pubmed-meshheading:16569439-Paramyxovirinae, pubmed-meshheading:16569439-Plasmids, pubmed-meshheading:16569439-RNA, Viral, pubmed-meshheading:16569439-Transfection, pubmed-meshheading:16569439-Vaccines, Attenuated, pubmed-meshheading:16569439-Vaccines, Synthetic, pubmed-meshheading:16569439-Vero Cells, pubmed-meshheading:16569439-Vesicular stomatitis Indiana virus, pubmed-meshheading:16569439-Viral Proteins, pubmed-meshheading:16569439-Viral Vaccines, pubmed-meshheading:16569439-Virus Replication
pubmed:year
2006
pubmed:articleTitle
An efficient helper-virus-free method for rescue of recombinant paramyxoviruses and rhadoviruses from a cell line suitable for vaccine development.
pubmed:affiliation
Wyeth Vaccines Research, 401 North Middletown Road, Pearl River, NY 10965, USA.
pubmed:publicationType
Journal Article