Goat corynebacterium pseudotuberculosis PLD recombinant protein as well as preparation method and application thereof

Abstract

The invention relates to goat corynebacterium pseudotuberculosis PLD recombinant protein as well as a preparation method and an application thereof, and belongs to the technical field of gene engineering. According to the recombinant protein provided by the invention, a sequence of a goat corynebacterium pseudotuberculosis phospholipase D (PLD) that signal peptide is removed is obtained by virtueof a whole-gene synthesis method; the sequence is sub-cloned into a prokaryotic expression vector pET28a; IPTG induction is conducted, so that soluble expression protein is obtained; and two times ofpurification are implemented, so that the expression protein that purity is above 95% is obtained. Based upon western-blot analysis, it is indicated that the protein has bio-activity; and through a series of optimization, a complete set of ELISA detection kits are assembled. Through sensitivity and specificity analysis as well as serological testing and immunological prevention monitoring on goat,testing and monitoring results show that the kits provided by the invention are relatively good in accuracy, sensitivity and repeatability; and the kits are applicable to diagnosis and immunologicalmonitoring of infection caused by goat corynebacterium pseudotuberculosis.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a goat pseudotuberculosis PLD recombinant protein and a preparation method and application thereof. Background technique [0002] Corynebacterium pseudotuberculosis is a facultative intracellular parasite, which mainly infects sheep and goats. The disease it causes is called caseous lymphadenitis. In terms of clinical manifestations, its main feature is purulent lymph nodes. The disease is currently in It is widely prevalent in the world, and there are many reports of this disease in China. Goat pseudotuberculosis is an internationally recognized animal disease that is difficult to purify. The important reason why it is difficult to prevent and treat is that there is a thick and dense fibrous granulomatous cyst on the surface of the abscess. As a result, it is difficult for the medicine to penetrate into it through this layer of cyst and play a role, so th...

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Problems solved by technology

[0002] Corynebacterium pseudotuberculosis is a facultative intracellular parasite, which mainly infects sheep and goats. The disease it causes is called caseous lymphadenitis. In terms of clinical manifestations, its main feature is purulent lymph nodes. The disease is currently in It is widely prevalent in the world, and there are many reports of this disease in China. Goat pseudotuberculosis is an internationally recognized animal disease that is difficult to purify. The important reason why it is difficult to prevent and treat is that there is a thick and dense fibrous granulomatous cyst on the surface of the abscess. As a result, it is difficult for drugs to infiltrate into this layer of cysts to play a role, so the effect of systemic medication is not good, and the procedures for local medication are cumbersome and difficult to eradicate

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