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含SipD和LptD免疫原的重组嵌合蛋白的生物信息学设计及其对鼠伤寒沙门氏菌免疫原性的评估

Bioinformatics design of recombinant chimeric protein containing SipD and LptD immunogens and evaluation of its immunogenicity against Salmonella Typhimurium.

作者信息

Aghaie Seyed Mojtaba, Tabatabaei Mohammad, Nazarian Shahram

机构信息

Department of Pathobiology, Faculty of Veterinary Medicine, Shiraz University, Shiraz, Iran.

Department of Pathobiology, Faculty of Veterinary Medicine, Shiraz University, Shiraz, Iran.

出版信息

Microb Pathog. 2023 Feb;175:105959. doi: 10.1016/j.micpath.2022.105959. Epub 2022 Dec 26.

Abstract

The growing emergence of resistant bacteria is the current global concern for the humans and animals. Vaccination could be the desirable method to preventing such infectious diseases. Safe and effective vaccines are urgently needed to manage and prevent Salmonella contamination. Subunit vaccines are safe approaches for the protection against Salmonella spp. The bioinformatics methods were performed to determine the gene structure. Gene cassette (rLPSI) was ordered in pET28a (+), and cloned into E.coli BL21 (DE3), and the recombinant protein was expressed using IPTG (1 mM). Mice were immunized by subcutaneous administration of recombinant protein. Then, the mice were challenged by oral administration of 100LD of live S. Typhimurium. The Codon adaptation index of the chimeric gene was multiplied by 0.92. Validation results showed that >90% of residues lie in the desired or extra allowed area of the Ramachandran plot. The recombinant protein (65.9 kDa) was expressed in E.coli. Antibody titers in vaccinated mice were significantly different from those in the control groups. Recombinant protein immunization of the mice provided 90% and 70% protection against 10LD and 100LD of S. Typhimurium, respectively. In general, the results showed the high efficiency of rLPSI chimeric protein as a protective antigen against S. Typhimurium infection. The rLPSI chimeric protein could be an effective recombinant vaccine candidate against S. Typhimurium infection, but more research is needed.

摘要

耐药细菌的不断出现是当前全球人类和动物所关注的问题。疫苗接种可能是预防此类传染病的理想方法。迫切需要安全有效的疫苗来控制和预防沙门氏菌污染。亚单位疫苗是预防沙门氏菌属感染的安全方法。运用生物信息学方法确定基因结构。将基因盒(rLPSI)克隆到pET28a(+)载体中,并导入大肠杆菌BL21(DE3),使用IPTG(1 mM)表达重组蛋白。通过皮下注射重组蛋白对小鼠进行免疫。然后,通过口服100LD的活鼠伤寒沙门氏菌对小鼠进行攻毒。嵌合基因的密码子适应指数乘以0.92。验证结果表明,>90%的残基位于拉氏图的理想或额外允许区域。重组蛋白(65.9 kDa)在大肠杆菌中表达。接种疫苗小鼠的抗体滴度与对照组有显著差异。重组蛋白免疫小鼠分别对10LD和100LD的鼠伤寒沙门氏菌提供了90%和70%的保护。总体而言,结果表明rLPSI嵌合蛋白作为抗鼠伤寒沙门氏菌感染的保护性抗原具有高效性。rLPSI嵌合蛋白可能是一种有效的抗鼠伤寒沙门氏菌感染的重组疫苗候选物,但还需要更多的研究。

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