Synthesis of mumps virus nucleocapsid protein in yeast Pichia pastoris.
The expression of mumps virus nucleocapsid protein in yeast Pichia pastoris was investigated. Viral nucleocapsid proteins usually elicit a strong long-term humoral immune response in patients and experimental animals. Therefore, the detection of antibodies specific to mumps virus nucleoprotein can play an important role in immunoassays for mumps diagnosis. For producing a high-level of recombinant mumps virus nucleoprotein the expression system of yeast P. pastoris was employed. The recombinant nucleocapsid protein was purified by cesium chloride ultracentrifugation of yeast lysates. Electron microscopy of the purified recombinant nucleocapsid protein revealed a herring-bone structure similar to the one discovered in mammalian cells infected with mumps virus. The yield of purified nucleocapsid-like particles from P. pastoris constituted 2.1 mg per 1 g of wet biomass and was considerably higher in comparison to the other expression systems.
Slibinskas
Rimantas
R
Institute of Biotechnology, V Graiciuno 8, Vilnius LT-2028, Lithuania.
Zvirbliene
Aurelija
A
Gedvilaite
Alma
A
Samuel
Dhanraj
D
Jin
Li
L
Beard
Stuart
S
Staniulis
Juozas
J
Sasnauskas
Kestutis
K
D000818:Animals; D003001:Cloning, Molecular; D015966:Gene Expression Regulation, Fungal / Q000502:physiology; D007118:Immunoassay / Q000379:methods; D051379:Mice; D009109:Mumps virus / Q000737:chemistry* / Q000378:metabolism*; D010843:Pichia / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011487:Protein Conformation; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D014760:Viral Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry* / Q000276:immunology
D011994:Recombinant Proteins; D014760:Viral Fusion Proteins