Abstract
The coronavirus membrane (M) protein acts as a dominant immunogen and is a major player in virus assembly. In this study, we prepared two monoclonal antibodies (mAbs; 1C3 and 4C7) directed against the transmissible gastroenteritis virus (TGEV) M protein. The 1C3 and 4C7 mAbs both reacted with the native TGEV M protein in western blotting and immunofluorescence (IFA) assays. Two linear epitopes, 243YSTEART249 (1C3) and 243YSTEARTDNLSEQEKLLHMV262 (4C7), were identified in the endodomain of the TGEV M protein. The 1C3 mAb can be used for the detection of the TGEV M protein in different assays. An IFA method for the detection of TGEV M protein was optimized using mAb 1C3. Furthermore, the ability of the epitope identified in this study to stimulate antibody production was also evaluated. An immunodominant epitope in the TGEV membrane protein endodomain was identified. The results of this study have implications for further research on TGEV replication.
Highlights
Coronaviruses (CoVs) are clustered in the Coronavirinae subfamily and are divided into four genera [1,2]
The recombinant proteins were expressed by induction with 1 mM IPTG in pCold-Transmissible gastroenteritis virus (TGEV)-M-transformed cells
Some immunodominant epitopes have been identified on the M proteins of the porcine epidemic diarrhoea virus (PEDV) [42], infectious bronchitis virus (IBV) [43] and severe acute respiratory syndrome (SARS)-CoV [44]
Summary
Coronaviruses (CoVs) are clustered in the Coronavirinae subfamily and are divided into four genera (alpha-, beta-, gamma-, and deltacoronavirus) [1,2]. CoVs are enveloped, single-stranded, positive-sense RNA viruses [3,4,5]. The CoV genomes range from 26.2 kb to 31.7 kb in size. Four structural proteins are encoded by the CoV genomes: spike (S), membrane (M), envelope (E), and nucleocapsid (N). Transmissible gastroenteritis virus (TGEV) is an excellent model of CoV biology [6,7,8,9,10,11,12]. The M protein is the viral assembly scaffold and the most abundant protein in the viral envelope [13]
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