1. [Expression of structural and non-structural proteins of severe fever with thrombocytopenia syndrome bunyavirus].
- Author
-
Lu J, Li C, Zhang FS, Wu W, Zhang QF, Zhang L, Wang T, Wang Q, Qiu PH, Liang MF, and Li DX
- Subjects
- Cell Line, Transformed, HEK293 Cells, Humans, Orthobunyavirus metabolism, Viral Nonstructural Proteins genetics, Viral Structural Proteins genetics, Virion metabolism, Bunyaviridae Infections virology, Fever virology, Orthobunyavirus genetics, Thrombocytopenia virology, Viral Nonstructural Proteins biosynthesis, Viral Structural Proteins biosynthesis, Virion genetics
- Abstract
Severe fever with thrombocytopenia syndrome bunyavirus (SFTSV) is a novel phlebovirus, causing a life-threatening illness associated with the symptoms of severe fever and thrombocytopenia syndrome. The sequence and structure of the genome have already been illustrated in previous study. However, the characteristics and function of the structure and non-structure proteins is still unclear. In this study, we identified the density of the purified SFTSV virions as 1.135 g/mL in sucrose solution. Using RT-PCR method, we amplified the full coding sequence of RNA dependent RNA polymerase(RdRp), glycoprotein precursor (M), glycoprotein n (Gn), glycoprotein c (Gc), nuclear protein (NP) and non structural protein (NSs) of SFTSV (strain HB29). Respectively inserted the target genes into eukaryotic expression vector pcDNA5/FRT or VR1012, the target protein in 293T cell were successfully expressed. By analyzing the SFTSV virions in SDS-PAGE and using recombinant viral proteins with SFTS patients sera in Western blotting and Immunofluorescent assay, the molecule weight of structure and non-structure proteins of SFTSV were defined. The study provides the first step to understand the molecular characteristics of SFTSV.
- Published
- 2011