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三元体自传媒粘合物SADA来自沙门氏菌spp. 作为一种新的细菌表面显示系统

Shuli Sang¹, Wenge Song¹, Lu Lu¹

¹State Key Laboratory of Pathogen and Biosecurity, Institute of Biotechnology, Academy of Military Medical Sciences, 20 Dongda Street, Fengtai District, Beijing 100071, China.

|April 27, 2024

概括

这项研究表明,使用沙门氏菌的V型分泌系统蛋白 SadA 来显示细菌表面上的外来蛋白质. 这种由SadB增强的细菌表面显示平台,显示了开发减弱细菌疫苗的潜力.

科学领域:

微生物学和分子生物学
细菌表面显示系统
疫苗开发疫苗开发

背景情况:

细菌表面显示平台用于疫苗输送和检测.
类型V分泌系统,如沙门氏菌中的SADA,有效地将外来蛋白质固定在格兰氏阴性细菌上.
SadA促进生物膜的形成和宿主细胞的粘附,而SadB促进其转移.

研究的目的:

为了研究SadaA作为一种定动机的有效性,在Salmonella typhimurium中进行异质蛋白表面表达.
评估SadB在增强SadA介导蛋白质显示中的作用.
评估基于SADA的显示系统在疫苗开发中的潜力.

主要方法:

使用SadaA及其截断的变种 (SadA^877,SadA^269,SadA^1292) 作为在沙门氏杆菌 typhimurium中的定图案.
与其内膜脂蛋白 SadB 共同表达的 SadA,以评估其对蛋白质显示的影响.
将异质蛋白 (mScarlet,尿酶B片段,PAD4) 移植到SADA结构上.
使用全细胞点斑,免疫光和流细胞计验证了表面定位.
在口服注射工程细菌后,评估体内免疫反应.

主要成果:

萨达A与萨达B结合,显著增强了沙门氏甲型菌表面的异质蛋白显示 (大约增加了三倍).
截断的SadaA变种 (SadA^877,SadA^269) 有效地显示了Helicobacter pylori尿酶B的B细胞表位.
较大的蛋白质结构,包括mScarlet和尿酶B-mScarlet融合,使用截断的SADA成功显示.
最大的构造,UreBm-PAD4-mScarlet,使用全长的 SadA^1292.2. 显示.
口服工程Salmonella菌株引发了显著的粘膜和幽默免疫反应.

结论:

在沙门氏菌中,SadaA作为一种有效的定基因,用于沙门氏菌中各种异质蛋白的表面表达.
在SadB的存在提高了SadaA介导的表面显示的效率.
基于SADA的细菌表面显示系统代表了开发减弱细菌疫苗的有希望的战略.

关键词:

悲伤的是悲伤的是悲伤沙门氏菌 spp. 沙门氏菌细菌表面显示显示器三重体的自传输体附着物,附着物和附着物.

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