非洲猪瘟病毒p30蛋白与CRM197的融合表达及对小鼠的免疫原性评价

doi:10.11843/j.issn.0366-6964.2024.11.038

Abstract

Abstract:

African swine fever is an acute, highly contagious and deadly infectious disease caused by the African swine fever virus, which has had a huge impact on the pig breeding industry. The lack of vaccines and effective therapeutic drugs has brought great challenges to the prevention and control of African swine fever. We tried to design a fusion protein to improve the immunogenicity of African swine fever virus p30. CRM197, a non-virulent mutant of diphtheria toxin, has been successfully used as a carrier protein in polysaccharide-protein conjugate vaccines to enhance the immunogenicity of polysaccharides. In this study, a fusion expression plasmid of p30 and CRM197 catalytic domain (A fragment) was constructed, and the immunogenicity of the purified recombinant protein and p30 alone were evaluated using mice. After immunizing mice with the fusion protein p30-CRM197(A) purified by prokaryotic expression and affinity chromatography, p30 alone could excite a high level of specific IgG against p30 in a shorter period of time, and maintain a high level of lymphocyte immune activity two months after the initial epidemic. The results of this study showed that the A fragment of CRM197 alone could significantly enhance the immunogenicity of prokaryotic expression p30 protein as a protein carrier, and it could be used as a candidate intramolecular adjuvant for African swine fever subunit vaccine.

Key words: African swine fever, CRM197, p30, immunogenicity

CLC Number:

S852.659.1

Qi ZHANG, Ningning DONG, Xiaomei TAN, Zhengwang SHI, Na LI, Wenqi ZHU, Aoxing TANG, Chuanfeng LI, Jie ZHU, Guangqing LIU, Yan SU, Chunchun MENG. Fusion Expression of African Swine Fever Virus p30 Protein and CRM197 and Evaluation of Its Immunogenicity in Mice[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(11): 5230-5237.

Figures/Tables 6

Table 1

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References 23

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目的基因 Target gene	寡核苷酸序列(5′→3′) Oligonucleotide sequence
p30	F: GCCATGGCTGATATCGGATCCATGAAAATGGAGGTCATC
p30	R: TCCACCACCACCAGATCCACCACCACCTTTTTTTTTTAAAAGTTTAATAAC
CRM197(A)	F: GGTGGTGGATCTGGTGGTGGTGGATCTATGGGCGCGGATGATGTG
CRM197(A)	R: CTCGAGTGCGGCCGCAAGCTTGCGCACGCGGTTGCCCGC

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Fusion Expression of African Swine Fever Virus p30 Protein and CRM197 and Evaluation of Its Immunogenicity in Mice

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