Construction of a mutagenesis cartridge for poliovirus genome-linked viral protein: isolation and characterization of viable and nonviable mutants
By following a strategy of genetic analysis of poliovirus, the authors 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 I (Mahoney) genome. The insertion of new restriction sites within the infectious clone has allowed them 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. They suggest that the position of the tyrosine residue is important for genome-linked viral protein function(s).
- Research Organization:
- State Univ. of New York (SUNY), Stony Brook, NY (United States)
- OSTI ID:
- 6244824
- Journal Information:
- Proc. Natl. Acad. Sci. U.S.A.; (United States), Vol. 85:2
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
POLIO VIRUS
MUTANTS
PROTEINS
AMINO ACID SEQUENCE
CODONS
ELECTROPHORESIS
HELA CELLS
MESSENGER-RNA
METHIONINE
MUTAGENESIS
OLIGONUCLEOTIDES
PHOSPHORUS 32
PLASMIDS
RECOMBINANT DNA
TRANSCRIPTION
TYROSINE
AMINO ACIDS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARBOXYLIC ACIDS
CELL CONSTITUENTS
DAYS LIVING RADIOISOTOPES
DNA
DRUGS
HYDROXY ACIDS
ISOTOPES
LIGHT NUCLEI
LIPOTROPIC FACTORS
MICROORGANISMS
MOLECULAR STRUCTURE
NUCLEI
NUCLEIC ACIDS
ODD-ODD NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PARASITES
PHOSPHORUS ISOTOPES
RADIOISOTOPES
RNA
VIRUSES
550201* - Biochemistry- Tracer Techniques