Construction and preparation of virus-like particle expression vector for yellow fever virus
10.3969/j.issn.1006-2483.2026.05.003
- VernacularTitle:黄热病毒的病毒样颗粒表达载体构建及制备
- Author:
Miaomiao LYV
1
;
Chunying GAO
1
;
Yunfeng LI
2
;
Zhiying JIN
2
;
Min WAN
2
;
Jie TIAN
2
;
Weina KANG
3
Author Information
1. School of Public Health, Hebei Medical University, Shijiazhuang, Hebei 050017, China;Science and Technology Research Center of China Customs, Beijing 100026, China
2. Science and Technology Research Center of China Customs, Beijing 100026, China
3. School of Public Health, Hebei Medical University, Shijiazhuang, Hebei 050017, China;Hebei Key Laboratory of Environment and Human Health, Shijiazhuang, Hebei 050017, China
- Publication Type:Journal Article
- Keywords:
Yellow fever virus;
Virus-like particles;
Expression vector construction;
Prokaryotic expression;
Nuclease resistance
- From:
Journal of Public Health and Preventive Medicine
2026;37(5):12-16
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare virus-like particles (VLPs) for yellow fever virus (YFV) using the pACYCDuet-1 single-plasmid dual expression system. Methods Gene fragments including NS1, NS5, and 5' untranslated region (UTR) and 3' UTR fragment of YFV were cloned into the expression plasmid pACYCDuet-1-MS2-TMpac (carrying MS2 coat protein gene) and transformed into E. coli BL21(DE3) for expression. VLPs were observed by TEM. After DNase I/RNase A digestion, RNA was extracted and transcribed into cDNA, which was subjected to fluorescence quantitative PCR detection. Results According to SN/T 3560-2013, double digestion and colony PCR of pACYCDuet-1-MS2-TMpac-YFV both produced expected bands. SDS-PAGE showed a target band at 25-35 kDa. RT-PCR amplified VLPs after enzyme digestion, and TEM revealed VLPs with clear edges and translucent interiors. Thus, the plasmid was successfully constructed, and the phage coat protein packaged YFV genes with nuclease resistance. Conclusion VLPs containing specific gene fragments of yellow fever virus have been successfully prepared. These nuclease-resistant VLPs can serve as safe and stable quality control materials to ensure the reliability of laboratory detection for YFV.
