[Expression of predicted B cell epitope peptide in S2 subunit of SARS coronavirus spike protein in E.coli and identification of its mimic antigenicity].
OBJECTIVE
To study the expression of predicted B cell epitope peptide in S2 subunit of SARS coronavirus spike protein in E.coli and its mimic antigenicity to S2 protein.


METHODS
B cell epitopes in S2 subunit of SARS coronavirus spike protein was predicted using DNAStar software. The cDNA sequence encoding the B cell epitope peptide was constructed artificially by PCR and then cloned into the downstream of chaperone 10 gene in vector pET28a(+) to construct pET28-chap10-S2epi plasmid. The fusion protein, chap10-S2epi, was expressed in E.coli BL21(DE3) and identified by SDS-PAGE and Western blot. The rabbit was immunized by purified Chap10-S2epi for the preparation of antiserum, which was used to identify the mimic antigenicity of Chap10-S2epi to S2 protein by ELISA.


RESULTS
Chap10-S2epi fusion protein was successfully constructed and expressed in E.coli. The antiserum from the animal immunized by Chap10-S2epi recognized full length of SARS coronavirus S2 spike protein.


CONCLUSIONS
The predicted B cell epitope peptide of SARS coronavirus S2 spike protein can induce the antigenicity of S2 protein, which provides some fundamental data for developing engineering vaccine against SARS coronavirus infection.
Feng
Yu
Y
Department of Molecular Biology, Basic Medical College, Jilin University, Changchun 130021, China.
Wan
Min
M
Wang
Xuan-jun
XJ
Zhang
Pei-yin
PY
Yu
Yong-li
YL
Wang
Li-ying
LY
D000818:Animals; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D018985:Epitopes, B-Lymphocyte / Q000235:genetics / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
D018985:Epitopes, B-Lymphocyte; D008562:Membrane Glycoproteins; D011993:Recombinant Fusion Proteins; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins; C578557:spike glycoprotein, SARS-CoV