Method for improving solubility of foot-and-mouth disease protein for immunization

Abstract

The present invention belongs to the technical field of preparation of a vaccine, and particularly relates to a method for improving the solubility of a foot-and-mouth disease protein for immunization. The method is by adding an His6-MBP protein tag in the C terminal of the foot-and-mouth disease protein by the mean genetic engineering in FMD protein by genetic engineering, so as to increase the solubility of the foot-and-mouth disease protein; the His6 represents the 6 histidine tags, and MBP represents a maltase binding protein. The present invention also provides an immune FMD protein CDG276 aiming at type A FMD prepared by the method. Compared to the FMDV VPI protein prepared by the existing genetic engineering, the FMD protein prepared by the invention has obvious increased the expression level; and due to the addition of soluble label maltose binding protein (MBP), the expressed FMDVA1 soluble protein has significantly increased solubility; therefore, the method has good application value.

Description

technical field [0001] The invention belongs to the technical field of vaccine preparation, and in particular relates to a method for improving the solubility of foot-and-mouth disease protein for immunity. Background technique [0002] Foot-and-mouth disease (FMD) is a severe contact infectious disease of cloven-hoofed animals caused by foot-and-mouth disease virus (FMDV), which mainly harms cattle, sheep, pigs, etc. It can cause a large-scale epidemic in the group and cause serious economic losses. The World Organization for Animal Health (OIE) classifies it as a notifiable disease, and the Ministry of Agriculture of my country also classifies it as a first-class animal infectious disease. The currently known FMDV includes A, O, C, Asia 1, SAT 1, SAT2 and SAT 3 serotypes without cross-immune protection, and more than 60 subtypes have been formed in the long-term mutual transmission infection. [0003] With the development of modern molecular biology technology, the resea...

AI Technical Summary

Problems solved by technology

Relatively speaking, the technical difficulties of protein expression and purification are relatively easy to overcome, while the soluble expression of proteins is a technical bottleneck

The cause and mechanism of protein insolubility are still in the exploratory stage, but it is generally believed that two factors lead to protein insolubility: one is the rate of protein translation, and the other is the rate of protein folding

[0005] In the existing research, one way to improve the solubility of protein expression is to improve the solubility of protein expression by expressing strains and changing the expression conditions, but the effect is sometimes not ideal, and due to the existence of inclusion bodies, the improvement effect of this way is unstable; another important and effective method is to add a soluble tag at the end of the active protein, through the fusion of the soluble tag and the target protein, and through the folding of the tag itself to increase the solubility of the target protein

However, different soluble tags can improve the solubility of different active proteins differently, and since soluble tags often need to be further removed in the later stage, it is often necessary to choose the most suitable soluble tag for different active proteins. Is the key, but also difficult work

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