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| Stable Expression of SARS-CoV-2 RBD Protein in Transgenic Rice |
| ZHANG Hui1, SONG Li1,*, ZHOU Yu2, HE Guo-Yu1, WANG Yue1, LI Qian-Feng2, PAN Zhi-Ming1,*, LIU Qiao-Quan2, JIAO Xin-An1 |
1 Jiangsu Key Laboratory of Zoonosis/Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses/Joint International Research Laboratory of Agriculture and Agri-product Safety of the Ministry of Education, Yangzhou University, Yangzhou 225009, China;
2 Key Laboratory of Plant Functional Genomics of the Ministry of Education/Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding, Yangzhou University, Yangzhou 225009, China |
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Abstract Since 2019, despite the rapid approval of many commercial vaccines, Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has continued to spread and mutate globally. The receptor binding domain (RBD) of the spike protein on the surface of SARS-CoV-2 is the main target for the development of COVID-19 (Coronavirus disease 2019) vaccines. To obtain a SARS-CoV-2 RBD protein that is low-cost, safe and easy to store and transport. The recombinant plant expression vector pCAMBIA1300-RBD was constructed by molecular cloning technique. Transgenic rice (Oryza sativa) plants of T0 generation were obtained by Agrobacterium tumefaciens-mediated rice genetic transformation. RBD gene in leaf genome of T0~T3 generation rice was detected by PCR, and the transcription and expression levels of RBD protein were identified by qRT-PCR and Western blot. The results showed that 2 homozygous T3 transgenic rice plants were successfully obtained, and RBD protein was stably expressed in rice seeds and could react specifically with Anti-S monoclonal antibody. The size of RBD protein treated by the peptide N-glycosidase F (PNGase F) was consistent with the expected size, indicating that the rice-derived RBD protein was N-glycosylated. In addition, the expression level of RBD protein in transgenic rice was determined by quantitative Western blot analysis. The results showed that the expression level of RBD protein in transgenic rice seeds was up to 312 μg/g in dry weight seeds. In summary, this study obtained homozygous transgenic plants stably expressing the SARS-CoV-2 RBD protein, providing a new approach for the further development of rice-derived subunit vaccines against Coronavirus Disease 2019 (COVID-19).
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Received: 06 January 2025
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Corresponding Authors:
*lisong@yzu.edu.cn; zmpan@yzu.edu.cn
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