Synthesis of the measles virus nucleoprotein in yeast Pichia pastoris and Saccharomyces cerevisiae.

Abstract	The development of a simple, efficient and cost-effective system for generation of measles virus nucleoprotein might help to upgrade reagents for measles serology. The gene encoding measles nucleoprotein was successfully expressed in two different yeast genera, Pichia pastoris and Saccharomyces cerevisiae, respectively. Both yeast genera synthesized a high level of nucleoprotein, up to 29 and 18% of total cell protein, in P. pastoris and S. cerevisiae, respectively. This protein is one of most abundantly expressed in yeast. After purification nucleocapsid-like particles (NLPs) derived from both yeast genera appeared to be similar to those detected in mammalian cells infected with measles virus. A spontaneous assembly of nucleoprotein into nucleocapsid-like particles in the absence of the viral leader RNA or viral proteins has been shown. Compartmentalisation of recombinant protein into large compact inclusions in the cytoplasm of yeast S. cerevisiae by green fluorescence protein (GFP) fusion has been demonstrated. Sera from measles patients reacted with the recombinant protein expressed in both yeast genera and a simple diagnostic assay to detect measles IgM could be designed on this basis.
Authors	Rimantas Slibinskas, Dhanraj Samuel, Alma Gedvilaite, Juozas Staniulis, Kestutis Sasnauskas
Journal	Journal of biotechnology (J Biotechnol) Vol. 107 Issue 2 Pg. 115-24 (Jan 22 2004) ISSN: 0168-1656 [Print] Netherlands
PMID	14711495 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)
Chemical References	Immunoglobulin M Luminescent Proteins Nucleoproteins Recombinant Fusion Proteins Green Fluorescent Proteins
Topics	Blotting, Western Cell Compartmentation Electrophoresis, Polyacrylamide Gel Escherichia coli Gene Dosage Green Fluorescent Proteins Humans Immunoglobulin M (blood) Kinetics Luminescent Proteins Measles (virology) Measles virus (chemistry, immunology, metabolism) Nucleoproteins (biosynthesis, genetics, immunology, isolation & purification, metabolism, ultrastructure) Pichia (metabolism) Recombinant Fusion Proteins (biosynthesis, metabolism) Saccharomyces cerevisiae (metabolism) Spectrometry, Fluorescence

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