TY - JOUR
T1 - Synthesis of recombinant human parainfluenza virus 1 and 3 nucleocapsid proteins in yeast Saccharomyces cerevisiae
AU - Juozapaitis, Mindaugas
AU - Zvirbliene, Aurelija
AU - Kucinskaite, Indre
AU - Sezaite, Indre
AU - Slibinskas, Rimantas
AU - Coiras, Mayte
AU - de Ory Manchon, Fernando
AU - López-Huertas, María Rosa
AU - Pérez-Breña, Pilar
AU - Staniulis, Juozas
AU - Narkeviciute, Irena
AU - Sasnauskas, Kestutis
PY - 2008/5
Y1 - 2008/5
N2 - Human parainfluenza virus types 1 and 3 (HPIV1 and HPIV3, respectively), members of the virus family Paramyxoviridae, are common causes of lower respiratory tract infections in infants, young children, the immunocompromised, the chronically ill, and the elderly. In order to synthesize recombinant HPIV1 and HPIV3 nucleocapsid proteins, the coding sequences were cloned into the yeast Saccharomyces cerevisiae expression vector pFGG3 under control of GAL7 promoter. A high level of recombinant virus nucleocapsid proteins expression (20-24 mg l-1 of yeast culture) was obtained. Electron microscopy demonstrated the assembly of typical herring-bone structures of purified recombinant nucleocapsid proteins, characteristic for other paramyxoviruses. These structures contained host RNA, which was resistant to RNase treatment. The nucleocapsid proteins were stable in yeast and were easily purified by caesium chloride gradient ultracentrifugation. Therefore, this system proved to be simple, efficient and cost-effective, suitable for high-level production of parainfluenza virus nucleocapsids as nucleocapsid-like particles. When used as coating antigens in an indirect ELISA, the recombinant N proteins reacted with sera of patients infected with HPIV1 or 3. Serological assays to detect HPIV-specific antibodies could be designed on this basis.
AB - Human parainfluenza virus types 1 and 3 (HPIV1 and HPIV3, respectively), members of the virus family Paramyxoviridae, are common causes of lower respiratory tract infections in infants, young children, the immunocompromised, the chronically ill, and the elderly. In order to synthesize recombinant HPIV1 and HPIV3 nucleocapsid proteins, the coding sequences were cloned into the yeast Saccharomyces cerevisiae expression vector pFGG3 under control of GAL7 promoter. A high level of recombinant virus nucleocapsid proteins expression (20-24 mg l-1 of yeast culture) was obtained. Electron microscopy demonstrated the assembly of typical herring-bone structures of purified recombinant nucleocapsid proteins, characteristic for other paramyxoviruses. These structures contained host RNA, which was resistant to RNase treatment. The nucleocapsid proteins were stable in yeast and were easily purified by caesium chloride gradient ultracentrifugation. Therefore, this system proved to be simple, efficient and cost-effective, suitable for high-level production of parainfluenza virus nucleocapsids as nucleocapsid-like particles. When used as coating antigens in an indirect ELISA, the recombinant N proteins reacted with sera of patients infected with HPIV1 or 3. Serological assays to detect HPIV-specific antibodies could be designed on this basis.
KW - ELISA
KW - Expression
KW - Human parainfluenza viruses (HPIV1, HPIV3)
KW - Nucleocapsid
KW - Nucleocapsid-like particles (NLPs)
KW - Yeast
UR - http://www.scopus.com/inward/record.url?scp=41049091345&partnerID=8YFLogxK
U2 - 10.1016/j.virusres.2007.12.016
DO - 10.1016/j.virusres.2007.12.016
M3 - Article
C2 - 18249456
AN - SCOPUS:41049091345
SN - 0168-1702
VL - 133
SP - 178
EP - 186
JO - Virus Research
JF - Virus Research
IS - 2
ER -