Expression of Poliovirus Complementary DNA Coding for Viral Antigenic Determinants in Escherichia coli1
- 1 August 1984
- journal article
- research article
- Published by Oxford University Press (OUP) in The Journal of Biochemistry
- Vol. 96 (2) , 305-312
- https://doi.org/10.1093/oxfordjournals.jbchem.a134839
Abstract
DNA sequences coding for the immunogenic capsid protein VPI and/or VP3 of poliovirus strain LSc-2ab (Sabin 1) were prepared by digesting the cloned complementary DNA with restriction endonuclease PstI . The DNA fragments were inserted into the unique PstI site of Escherichia coil plasmid vectors pBR322, pKT 280 and /or pKT 287 that lay in the region expressed under control of the penicillinase promoter system. In the expression vectors, poliovirus sequences were designed to be read in phase and therefore to be expressed as fusion proteins with the bacterial peptides. In addition, the Escherichia coil tryptophan operon promoter-operator system was inserted upstream of the penicillinase system to obtain stronger expression of the poliovirus sequences. Escherichia coil transformed with these plasmids appeared to produce the antigenic polypeptides, which were detected by immunoprecipitation with antibodies to capsid proteins VPI and /or VP3 followed by SDS-polyacrylamide gel electrophoresis.Keywords
This publication has 7 references indexed in Scilit:
- Complete nucleotide sequence of the attenuated poliovirus Sabin 1 strain genome.Proceedings of the National Academy of Sciences, 1982
- Restriction Map of Double-Stranded DNA Copy Synthesized from Poliovirus Sabin 1 RNA1The Journal of Biochemistry, 1982
- Molecular cloning of poliovirus cDNA and determination of the complete nucleotide sequence of the viral genome.Proceedings of the National Academy of Sciences, 1981
- Primary structure, gene organization and polypeptide expression of poliovirus RNANature, 1981
- Construction of plasmid vectors with unique PstI cloning sites in a signal sequence coding regionGene, 1980
- Changes in three of the four coat proteins of oral polio vaccine strain derived from type 1 poliovirusJournal of Virology, 1980
- Nucleotide sequence of the ampicillin resistance gene of Escherichia coli plasmid pBR322.Proceedings of the National Academy of Sciences, 1978