Construction of expression plasmids for Saccharomyces cerevisiae: application for synthesis of poliovirus protein VP2.
A series of expression plasmids was constructed to compare the usefulness of various promoters for the synthesis of a given protein in the Saccharomyces cerevisiae. The plasmids pMBL212, -213, -214, -215 and -216 can be used to synthesize the protein of interest directly as a non-fused protein or, if the protein is difficult to detect, indirectly as an enzymatically active beta-galactosidase fusion protein. The plasmids were employed to identify which yeast promoter and strain are suitable for the synthesis of poliovirus protein VP2. It was concluded that the GAL7 and PGK promoters in combination with strain X904 can be used for efficient synthesis of a VP2 in the form of a N-terminally fused VP2-beta-galactosidase protein.
D001483:Base Sequence; D002213:Capsid / Q000235:genetics*; D036022:Capsid Proteins; D004797:Enzyme-Linked Immunosorbent Assay; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000276:immunology; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D001616:beta-Galactosidase / Q000235:genetics
D036022:Capsid Proteins; D011993:Recombinant Fusion Proteins; C442541:VP2 protein, Poliovirus; D001616:beta-Galactosidase