Optimization Expression of Plant-derived SARS-CoV-2 S1-RBD
10.13865/j.cnki.cjbmb.2024.10.1225
- VernacularTitle:植物源SARS-CoV-2 S1-RBD表达水平的优化
- Author:
Han LI
1
;
Hui-Ying JI
;
Tian-Hao ZHANG
;
Chun-Yu WANG
;
Jin-Xin HE
;
Li-Li ZHANG
Author Information
1. 山西农业大学动物医学学院 动物医学系,山西 太谷030801;中国科学院微生物研究所 植物基因组学国家重点实验室,北京100101
- Publication Type:Journal Article
- Keywords:
SARS-CoV-2;
receptor-binding domain;
plant expression system;
gene silencing suppressors;
signal peptide;
immunogenicity
- From:
Chinese Journal of Biochemistry and Molecular Biology
2024;40(12):1761-1770
- CountryChina
- Language:Chinese
-
Abstract:
The receptor-binding domain (RBD) of SARS-CoV-2 is a key target for recombinant protein vaccines.Efficient,high-quality expression of RBD protein is crucial for developing effective recombinant protein vaccines.Compared to RBD vaccines produced in CHO cells,plant expression systems offer ad-vantages of higher safety and lower cost.However,current RBD antigen expression levels in plants are relatively low,and methods for extraction and quality optimization have not been fully established.This study used amino acids R319～K537 of the SARS-CoV-2 prototype strain RBD as a template for transient expression in Nicotiana benthamiana using the plant expression vector pCAMBIA1300.The initial expres-sion levels were low at 23.3 μg/g fresh weight,accounting for 0.23％ of total soluble plant protein.There was a 2.61-fold increase of RBD expression when it was co-expressed with the efficient gene silen-cing suppressor pGD-VSRs.By fusing green fluorescent protein to the C-terminus,we localized RBD ex-pression to the apoplastic region in plants.Further introduction of the αAmy3 signal peptide improved RBD accumulation with a 2.1-fold compared to the PR1a signal peptide.Ultimately,RBD expression in N.benthamiana was enhanced to 128.6μg/g fresh weight,constituting 1.29％of total soluble plant pro-tein.Subsequently,we employed an apoplastic extraction and purification method to obtain RBD protein with over 90％ purity.Mouse immunization experiments demonstrated that this plant-derived RBD has comparable immunogenicity to CHO-derived RBD.This study provides important theoretical foundation and technical support for the development of plant-derived RBD vaccines.
