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Preparation of virus-like particles of human GⅡ.4 norovirus in the flashBAC baculovirus expression system
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Zhipeng LIU1, Jinning XIAO2, Liangwei DUAN1, Qiongzi WANG1, Xiangpeng WANG1, *
Acta Microbiologica Sinica | 2024, 64(10) : 3798 - 3808
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Acta Microbiologica Sinica | 2024, 64(10): 3798-3808
• Research Articles •
Preparation of virus-like particles of human GⅡ.4 norovirus in the flashBAC baculovirus expression system
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Zhipeng LIU1, Jinning XIAO2, Liangwei DUAN1, Qiongzi WANG1, Xiangpeng WANG1, *
Affiliations
  • 1 Henan Key Laboratory of Immunology and Targeted Drugs, School of Medical Technology, Xinxiang Medical University, Xinxiang 453003, Henan, China
  • 2 Department of Clinical Laboratory, the Third Affiliated Hospital of Xinxiang Medical University, Xinxiang 453003, Henan, China
Published: 2024-06-21 doi: 10.13343/j.cnki.wsxb.20240206
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[Objective] Human norovirus (HuNoV) is one of the most common pathogens causing acute gastroenteritis in humans worldwide. Currently, there are no approved vaccines to prevent this disease. This study aimed to prepare virus-like particles (VLPs) of HuNoV in the flashBAC baculovirus expression system, laying a foundation for the development of vaccines against HuNoV. [Methods] After codon optimization, the full-length gene sequence of the VP1 protein of HuNoV GⅡ.4 was synthesized and cloned into the baculovirus pBacPAK9 transfer vector to obtain the recombinant plasmid pBacPAK9-8his-VP1. After enzyme digestion and sequencing, the recombinant plasmid was co-transfected with the linear baculovirus plasmid (Bacmid) into SF9 cells to obtain a recombinant baculovirus carrying the VP1 gene. Hi-Five (HF) cells were infected by the recombinant baculovirus for protein expression, and the expression of VP1 was analyzed by SDS-PAGE and Western blotting. VP1 was purified by Ni-NTA affinity chromatography and identified by SDS-PAGE and Western blotting. The purity of VP1 was examined by high performance liquid chromatography (HPLC). The VLPs were observed by transmission electron microscopy. [Results] A transfer plasmid pBacPAK9-8his-VP1 was constructed, and the VP1 protein, with a molecular weight of approximately 58 kDa, was mainly expressed in the cytoplasm of HF cells. The HPLC results showed that the purity of VP1 was over 99%. The VLPs with a regular shape, uniform sizes, and diameters of 30–40 nm were observed by transmission electron microscopy. [Conclusion] The VLPs of HuNoV GⅡ.4 were prepared with a baculovirus expression system, laying a foundation for the development of HuNoV vaccines.

human norovirus  /  VP1 protein  /  baculovirus expression system  /  virus-like particles
Zhipeng LIU, Jinning XIAO, Liangwei DUAN, Qiongzi WANG, Xiangpeng WANG. Preparation of virus-like particles of human GⅡ.4 norovirus in the flashBAC baculovirus expression system[J]. Acta Microbiologica Sinica, 2024 , 64 (10) : 3798 -3808 . DOI: 10.13343/j.cnki.wsxb.20240206
Funding
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  • Key Scientific and Technological Project of Henan Province(232102311062)
  • Key Scientific and Technological Project of Henan Province(222102310688)
Appendix
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Year 2024 volume 64 Issue 10
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110
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Article Info
doi: 10.13343/j.cnki.wsxb.20240206
  • Receive Date：2024-03-29
  • Online Date：2026-03-21
  • Published：2024-06-21
Article Data
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History
  • Received：2024-03-29
  • Accepted：2024-06-17
Funding
Key Scientific and Technological Project of Henan Province(232102311062)
Key Scientific and Technological Project of Henan Province(222102310688)
Affiliations
    1 Henan Key Laboratory of Immunology and Targeted Drugs, School of Medical Technology, Xinxiang Medical University, Xinxiang 453003, Henan, China
    2 Department of Clinical Laboratory, the Third Affiliated Hospital of Xinxiang Medical University, Xinxiang 453003, Henan, China

Corresponding:

*WANG Xiangpeng, E-mail:
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表12种不同金属材料的力学参数
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鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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