Intracellular location of severe fever with thrombocytopenia syndrome bunyavirus NP protein with correlative light and electron microscopy based on miniSOG
10.3760/cma.j.issn.1003-9279.2018.01.016
- VernacularTitle: 利用光镜-电镜关联方法研究发热伴血小板减少综合征病毒核蛋白的亚细胞定位
- Author:
Guoyu NIU
1
;
Zun ZHANG
2
,
3
;
Baicheng XIA
1
;
Shuhui LIU
1
;
Xu GAO
1
;
Wei WU
2
;
Xiaohui ZOU
2
;
Zhuozhuang LU
2
;
Tao HONG
2
Author Information
1. School of Public Health and Management, Weifang Medical University, Weifang 261053, China
2. National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China
3. Henan Chemical Technician College, Kaifeng 475000, China
- Publication Type:Journal Article
- Keywords:
Mini singlet oxygen generator;
Severe fever with thrombocytopenia syndrome virus;
Nucleoprotein;
Correlative light and electron microscopy
- From:
Chinese Journal of Experimental and Clinical Virology
2018;32(1):75-79
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To study the intracellular location and characteristic of SFTSV NP protein in different phases using mini singlet oxygen generator (miniSOG) labeling technique.
Methods:MiniSOG is a recently-invented genetically-encoded tag for EM. MiniSOG-fused SFTSV NP (NPSOG) gene was cloned by PCR, and inserted into pcDNA3.0 plasmid to form pTPL-NPSOG, which was used to transfect 293 cells. The transfected cells of different phases were fixed in 2.5% glutaraldehyde in situ, stained with DAB through the photooxidation activity of miniSOG, and used to prepare ultrathin sections. Intracellular location and characteristic of SFTSV NP protein in different phases were studied by observing the sections under transmission electron microscope.
Results:After transfecting the plasmid with NPSOG to 293 cells, NP protein was expressed in cytoplasm and peri nucleus, and gradually aggregated, which connected with endoplasmic reticulum and Golgi apparatus to form larger volume and irregular inclusion bodies in cytoplasm. No obvious subcellular structure changes were found.
Conclusions:The SFTSV nucleoprotein can be expressed separately to form inclusion bodies without the assistance of other viral proteins. The formation of inclusion bodies requires the directional movement and aggregation of a certain number of NP proteins, which may involve the interaction of NP protein and host organelles during this period.
