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.的ROP27蛋白的免疫保护作用

The Immunoprotective Effect of ROP27 Protein of .

作者信息

Li Menggang, Lv Xiaoling, Zheng Mingxue, Wei Yingyi

机构信息

Guangxi Key Laboratory of Animal Breeding, Disease Control and Prevention, College of Animal Science and Technology, Nanning 530004, China.

College of Veterinary Medicine, Shanxi Agricultural University, Jinzhong 030801, China.

出版信息

Animals (Basel). 2023 Nov 13;13(22):3500. doi: 10.3390/ani13223500.

Abstract

rhoptry protein has the properties of a protective antigen. ROP27 is a pathogenic gene that is detected via a transcriptome, but its expression pattern, immunogenicity, and potency are unknown. Therefore, a gene segment of ROP27 was amplified and transplanted into the pET28a prokaryotic vector for the expression of the recombinant protein, and it subsequently purified for the generation of a polyclonal antibody. Then, RT-PCR and Western blotting were performed to understand the expression pattern of ROP27. Subsequently, animal experiments were conducted to evaluate the immunoprotective effect of the recombinant protein with different immunizing doses (50, 100, and 150 μg). The results showed that the expression of ROP27 gradually increased with the prolongation of infection time, reaching the highest level at 96 h and then decreasing. Additionally, ROP27 is a natural antigen of coccidia that can stimulate the body to produce high levels of IgY. As with recombinant protein vaccines, the results of immune protection evaluation tests showed that the average weight gain rates of the immune challenge groups were significantly higher than that of the challenged control group, and their average lesion scores were significantly lower than that of the challenged control group. Furthermore, the oocyst excretion decreased by 81.25%, 86.21%, and 80.01%, and the anticoccidial index was 159.45, 171.47, and 166.75, respectively, for these groups. ROP27 is a promising antigen gene candidate for the development of a coccidiosis vaccine.

摘要

棒状体蛋白具有保护性抗原的特性。ROP27是一种通过转录组检测到的致病基因,但其表达模式、免疫原性和效力尚不清楚。因此,扩增了ROP27的基因片段并将其克隆到pET28a原核载体中以表达重组蛋白,随后对其进行纯化以制备多克隆抗体。然后,进行RT-PCR和蛋白质印迹分析以了解ROP27的表达模式。随后,进行动物实验以评估不同免疫剂量(50、100和150μg)的重组蛋白的免疫保护效果。结果表明,ROP27的表达随着感染时间的延长而逐渐增加,在96小时达到最高水平,然后下降。此外,ROP27是球虫的天然抗原,可刺激机体产生高水平的IgY。与重组蛋白疫苗一样,免疫保护评估试验结果表明,免疫攻毒组的平均体重增加率显著高于攻毒对照组,其平均病变评分显著低于攻毒对照组。此外,这些组的卵囊排泄分别减少了81.25%、86.21%和80.01%,抗球虫指数分别为159.45、171.47和166.75。ROP27是开发球虫病疫苗的一个有前景的抗原基因候选物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5097/10668730/02c150d53c63/animals-13-03500-g001.jpg

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