Vetráková A, Chovanová R Kalianková, Rechtoríková R, Krajčíková D, Barák I
Department of Microbial Genetics, Institute of Molecular Biology, Slovak Academy of Sciences, Bratislava, Slovakia.
Comput Struct Biotechnol J. 2023;21:1550-1556. doi: 10.1016/j.csbj.2023.02.007. Epub 2023 Feb 8.
spores are considered to be efficient and useful vehicles for the surface display and delivery of heterologous proteins. In this study, we prepared recombinant spores with the receptor binding domain (RBD) of the SARS-CoV-2 spike glycoprotein displayed on their surface in fusion with the CotZ or CotY spore coat proteins as a possible tool for the development of an oral vaccine against the SARS-CoV-2 virus. The RBD was attached to the N-terminus or C-terminus of the coat proteins. We also directly adsorbed non-recombinantly produced RBD to the spore surface. SDS-PAGE, western blot and fluorescence microscopy were used to analyze RBD surface expression on purified spores. Results obtained from both display systems, recombinant and non-recombinant, demonstrated that RBD was present on the spore surfaces.
孢子被认为是用于异源蛋白表面展示和递送的有效且有用的载体。在本研究中,我们制备了重组孢子,其表面展示有严重急性呼吸综合征冠状病毒2(SARS-CoV-2)刺突糖蛋白的受体结合域(RBD),该RBD与CotZ或CotY孢子壁蛋白融合,作为开发针对SARS-CoV-2病毒的口服疫苗的一种可能工具。RBD连接在壁蛋白的N端或C端。我们还将非重组产生的RBD直接吸附到孢子表面。使用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)、蛋白质免疫印迹法(western blot)和荧光显微镜来分析纯化孢子上RBD的表面表达。从重组和非重组这两种展示系统获得的结果表明,RBD存在于孢子表面。