Sheep aphthosis virus recombinant subunit vaccine, and production method thereof

Abstract

The invention relates to a sheep aphthosis virus fusion protein rB2LcF1L. An amino acid sequence of the sheep aphthosis virus B2L and F1L fusion protein is SEQ ID No:2; compared with the wild type sheep aphthous virus F1L protein, the sheep aphthosis virus fusion protein rB2LcF1L has amino acid sequences of sites No.1 to 70 deleted to facilitate the expression of the fusion protein and increase the expression level. At the same time, 6 groups of histidine tags are added to the C-terminus of the fusion protein to facilitate purification in subsequent production. In the above amino acid sequence, compared with a B2L amino acid sequence of a conventional isolate, the V / L of the amino acid at a site No.9 is mutated to G, namely ''IPVG or IPLG is mutated to IPGG'', and the LDCF at a site No.35is mutated to LYCF. The above mutation significantly increases the expression level and immunogenicity of the fusion protein.

Description

Technical field: [0001] The invention belongs to the field of veterinary biological products, and in particular relates to a recombinant subunit vaccine of aphthous ulcer virus and a production method thereof. Background technique: [0002] Oral sores, also known as sheep infectious pustule, is a contact and epithelial infectious disease that seriously threatens the sheep industry. It mainly harms 3-6 month-old lambs and some adult sheep. The clinical symptoms of the disease are skin rashes, pustules, ulcers and scabs on the lips and mucous membranes of the mouth. Lambs show ulcers on the teeth and lips and cannot eat. The incidence rate is as high as 100%. If secondary or mixed infection with other pathogens Microbial mortality is high, up to 15%. Mouth occurs worldwide and is widespread. In recent years, with the development of my country's sheep raising industry and the change of feeding methods, the risk of the disease has increased, the affected area and the severity ...

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Problems solved by technology

However, like the literature report by Liu Yuan et al., the method of the patent application for this invention can only achieve transient expression in eukaryotic cells, and cannot obtain a stably expressed subunit vaccine, and involves the transformation of multiple plasmids. Plasmid loss is also prone to occur in , which cannot obtain genetically stable expressing cell lines

[0005] Based on the above technical research, it is shown that the current oral ulcer virus is difficult to culture and proliferate on conventional cell lines, and is not suitable for making inactivated vaccines or attenuated vaccines, and the existing research on subunit vaccines or nucleic acid vaccines also has certain defects, for example, genetic Poor stability, unable to obtain genetically stable expression cell lines, poor immunogenicity or low protection rate, etc.

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