Location of Repository

Construction of a "mutagenesis cartridge" for poliovirus genome-linked viral protein: isolation and characterization of viable and nonviable mutants.

By R J Kuhn, H Tada, M F Ypma-Wong, J J Dunn, B L Semler and E Wimmer

Abstract

By following a strategy of genetic analysis of poliovirus, we have constructed a synthetic "mutagenesis cartridge" spanning the genome-linked viral protein coding region and flanking cleavage sites in an infectious cDNA clone of the type 1 (Mahoney) genome. The insertion of new restriction sites within the infectious clone has allowed us to replace the wild-type sequences with short complementary pairs of synthetic oligonucleotides containing various mutations. A set of mutations have been made that create methionine codons within the genome-linked viral protein region. The resulting viruses have growth characteristics similar to wild type. Experiments that led to an alteration of the tyrosine residue responsible for the linkage to RNA have resulted in nonviable virus. In one mutant, proteolytic processing assayed in vitro appeared unimpaired by the mutation. We suggest that the position of the tyrosine residue is important for genome-linked viral protein function(s)

Topics: Research Article
Year: 1988
OAI identifier: oai:pubmedcentral.nih.gov:279582
Provided by: PubMed Central
Sorry, our data provider has not provided any external links therefor we are unable to provide a PDF.

Suggested articles


To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.