徐本锦;宣焱;杜淼;刘玲;吴惠文;宋彬妤;侯艳香;陈利荣;刘晓梁;

• 1:山西医科大学汾阳学院医学检验系
• 2:山西医科大学汾阳学院科技中心

摘要(Abstract)：

SARS-CoV-2是一种高致病性且传播迅速的病原体,通过刺突糖蛋白(Spike glycoprotein,S蛋白)识别宿主细胞表面的受体来实现入侵和感染。对S蛋白进行系统的生物信息学分析和原核表达,有助于深入理解S蛋白的功能和阐明该蛋白介导病毒感染的分子机制。本文采用Protparam、Pfam、TMHMM、ExPASy-ProtScale、PSORTⅡ、SignalP、UniProt、NetPhos 3.1、NetNGlyc 1.0、NetOGlyc 4.0和BLAST等生物信息学软件和数据库对S蛋白的理化性质、亚细胞定位、翻译后修饰及相互作用网络等生物学特性进行了系统分析。利用Clustal X2和MEGA7.0软件对该蛋白进行了基于氨基酸序列的同源性分析和系统进化分析。最后,通过分子克隆技术构建重组表达载体pET-22b-S并进行原核表达。结果显示,S蛋白由1 273个氨基酸组成,分子量141.2 kD,等电点6.24,有两个卷曲螺旋结构,一个跨膜螺旋区,疏水性较强。S蛋白包含刺突受体结合结构域和S2糖蛋白结构域,主要分布于宿主细胞的内质网膜和细胞膜,含有136个潜在的磷酸化位点和20个可能的糖基化位点。与SARS-CoV-2 S蛋白序列一致性最高的是SARS冠状病毒、SARS冠状病毒WH20和蝙蝠冠状病毒HKU3,均为76%。SARS-CoV-2与SARS冠状病毒和蝙蝠冠状病毒聚为一大支,提示它们可能具有共同的祖先。S蛋白主要在细菌裂解液离心之后的沉淀中表达,这为后续的结构分析和疫苗研发奠定了基础。S蛋白在SARS冠状病毒和蝙蝠冠状病毒之间保守性较高,提示其在病毒入侵过程中具有重要功能。SARS-CoV-2与SARS冠状病毒和蝙蝠冠状病毒可能具有共同的祖先。本研究为SARS-CoV-2 S蛋白的表达纯化、结构与功能研究提供了重要的数据基础,有助于全面揭示S蛋白的生物学功能,同时为设计和筛选靶向S蛋白的新型抗病毒药物提供了科学依据。

关键词(KeyWords)： 新冠病毒(SARS-CoV-2);刺突糖蛋白(S蛋白);生物信息学分析;原核表达

Abstract:

Keywords:

基金项目(Foundation): 山西医科大学汾阳学院引进人才启动金项目（项目号：2020A01），题目：RelA介导大肠杆菌严谨反应的分子机制研究;; 山西省高等学校科技创新项目（项目号：2020L0749），题目：大肠杆菌核糖体再循环因子RRF的功能研究;; 国家级大学生创新创业训练计划项目（项目号：202117114001），题目：新冠病毒Nsp1基因的克隆、表达载体构建及其对大肠杆菌生长的影响;; 山西省高等学校大学生创新创业训练计划重点项目（项目号：S202117114008），题目：SARS-CoV-2核衣壳蛋白过表达影响细菌蛋白翻译的分子机制;; 吕梁市科技计划项目（项目号：2020SHFZ29），题目：吕梁市食源性“超级耐药菌”MRSA的遗传多态性与溯源研究;吕梁市科技计划项目（项目号：2020XGZX104），题目：新冠肺炎疾病转归预测和风险评估的模型构建与评估~~

作者(Authors): 徐本锦;宣焱;杜淼;刘玲;吴惠文;宋彬妤;侯艳香;陈利荣;刘晓梁;

DOI: 10.13242/j.cnki.bingduxuebao.004090

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