[Preparation and characterization of the epitopes of SARS coronavirus spike protein].
OBJECTIVE
To prepare the recombinant epitopes of SARS-CoV spike protein and study their antigenic property to spike protein.


METHODS
The epitopes of SARS-CoV spike protein were analyzed by epitope prediction software. A novel gene, named Z, encoding 16 predicted epitopes of spike protein was designed and synthesized using chemical method. Z gene was cloned into vector pET-28a(+), expressed in E.coli BL21(DE3) and purified by Ni(2+) affinity method. Z protein was used as antigen to immunize the rabbit and anti-Z sera was collected. Then the antigenic property of Z protein to SARS-CoV spike protein was analyzed by dot-blot and ELISA assay.


RESULTS
Z gene was successfully designed and expressed in E.coli BL21(DE3). Dot blot analysis showed that SARS-CoV spike protein, which was expressed and purified from mammal cells, can be detected by anti-Z sera from rabbit. ELISA analysis indicated the SARS-CoV antigen prepared from SARS-CoV lysates can be detected by anti-Z sera.


CONCLUSIONS
The predicted epitopes of Z protein can induce the development of SARS-CoV spike protein antibody in rabbits, which provides a new protein for developing vaccine against SARS-CoV.
Fang
Kai
K
Department of Bioinformatics, College of Basic Medical Sciences, China Medical University, Shenyang 110001, China.
Zhong
Lian-Sheng
LS
Ma
Ru-Hai
RH
Wang
Shao-Cheng
SC
Pan
Zhong-Cheng
ZC
Zhang
Yu-Kui
YK
He
Qun
Q
Zhao
Yu-Jie
YJ
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D019295:Computational Biology; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000032:analysis / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics; D007106:Immune Sera / Q000032:analysis / Q000276:immunology; D008562:Membrane Glycoproteins / Q000276:immunology*; D008969:Molecular Sequence Data; D045473:Severe acute respiratory syndrome-related coronavirus / Q000276:immunology*; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins / Q000276:immunology*
D000939:Epitopes; D007106:Immune Sera; D008562:Membrane Glycoproteins; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins; C578557:spike glycoprotein, SARS-CoV