Efficient Production of Enterovirus 71 (EV71) Virus-like Particles by Controlling Promoter Strength in Insect Cells

被引:3
作者
Kim, Hyun-Soo [1 ]
Moon, Hyuk-Jin [1 ]
Choi, Jae-Bang [2 ]
Han, Beom-Ku [2 ]
Woo, Soo Dong [1 ]
机构
[1] Chungbuk Natl Univ, Coll Agr Life & Environm Sci, Dept Agr Biol, Cheongju 28644, South Korea
[2] Optipharm Inc, Osong 28158, South Korea
来源
VIRUSES-BASEL | 2024年 / 16卷 / 06期
关键词
HFMD; enterovirus; 71; VLP; baculovirus; burst sequences; BACULOVIRUS EXPRESSION VECTOR; PURIFICATION;
D O I
10.3390/v16060834
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
This study was conducted to efficiently produce virus-like particles (VLPs) of enterovirus 71 (EV71), a causative virus of hand, foot, and mouth disease (HFMD). The expression level of the P1 precursor, a structural protein of EV71, was modified to increase VLP production, and the optimal expression level and duration of the 3CD protein for P1 cleavage were determined. The expression level and duration of 3CD were controlled by the p10 promoter, which was weakened by repeated burst sequence (BS) applications, as well as the OpIE2 promoter, which was weakened by the insertion of random untranslated region sequences of various lengths. The cleavage and production efficiency of the P1 precursor were compared based on the expression time and level of 3CD, revealing that the p10-BS5 promoter with four repeated BSs was the most effective. When P1 and 3CD were expressed using the hyperexpression vector and the p10-BS5 promoter, high levels of structural protein production and normal HFMD-VLP formation were observed, respectively. This study suggests that the production efficiency of HFMD-VLPs can be significantly enhanced by increasing the expression of the P1 precursor and controlling the amount and duration of 3CD expression.
引用
收藏
页数:11
相关论文
共 26 条
[1]   Vaccine delivery: a matter of size, geometry, kinetics and molecular patterns [J].
Bachmann, Martin F. ;
Jennings, Gary T. .
NATURE REVIEWS IMMUNOLOGY, 2010, 10 (11) :787-796
[2]   Utility of Alternative Promoters for Foreign Gene Expression Using the Baculovirus Expression Vector System [J].
Bruder, Mark R. R. ;
Aucoin, Marc G. G. .
VIRUSES-BASEL, 2022, 14 (12)
[3]   Clinical characteristics and managements of severe hand, foot and mouth disease caused by enterovirus A71 and coxsackievirus A16 in Shanghai, China [J].
Cai, Kang ;
Wang, Yizhong ;
Guo, Zhongqin ;
Yu, Huiju ;
Li, Huajun ;
Zhang, Liya ;
Xu, Shanshan ;
Zhang, Qingli .
BMC INFECTIOUS DISEASES, 2019, 19 (1)
[4]   Determination of the cleavage site of enterovirus 71 VP0 and the effect of this cleavage on viral infectivity and assembly [J].
Cao, Jiaming ;
Liu, Hongtao ;
Qu, Meng ;
Hou, Ali ;
Zhou, Yan ;
Sun, Bo ;
Cai, Linjun ;
Gao, Feng ;
Su, Weiheng ;
Jiang, Chunlai .
MICROBIAL PATHOGENESIS, 2019, 134
[5]  
Chong P.C.S., 2018, Plotkins Vaccines, V7th ed., P288, DOI [10.1016/B978-0-323-35761-6.00021-3, DOI 10.1016/B978-0-323-35761-6.00021-3]
[6]   Enterovirus 71 virus-like particle vaccine: Improved production conditions for enhanced yield [J].
Chung, Cheng-Yu ;
Chen, Chi-Yuan ;
Lin, Shih-Yeh ;
Chung, Yao-Chi ;
Chiu, Hsin-Yi ;
Chi, Wei-Kuang ;
Lin, Yu-Li ;
Chiang, Bor-Luen ;
Chen, Wei-Jheng ;
Hu, Yu-Chen .
VACCINE, 2010, 28 (43) :6951-6957
[7]   Expression, purification and characterization of enterovirus-71 virus-like particles [J].
Chung, Yao-Chi ;
Huang, Jen-Huang ;
Lai, Chia-Wei ;
Sheng, Heng-Chun ;
Shih, Shin-Ru ;
Ho, Mei-Shang ;
Hu, Yu-Chen .
WORLD JOURNAL OF GASTROENTEROLOGY, 2006, 12 (06) :921-927
[8]   Virus-Like Particle Engineering: From Rational Design to Versatile Applications [J].
Ding, Xuanwei ;
Liu, Dong ;
Booth, George ;
Gao, Wei ;
Lu, Yuan .
BIOTECHNOLOGY JOURNAL, 2018, 13 (05)
[9]  
Gopal R, 2018, METHODS MOL BIOL, V1776, P125, DOI 10.1007/978-1-4939-7808-3_8
[10]   Development of a novel enhanced baculovirus expression vector via promoter combination [J].
Gwak, Won-Seok ;
Kim, Hyun-Soo ;
Bae, Jun-Su ;
Kim, Tae-Hee ;
Bae, Sung-Min ;
Woo, Soo-Dong .
JOURNAL OF ASIA-PACIFIC ENTOMOLOGY, 2020, 23 (04) :909-914