Antiserum Levels in Mice Inoculated with Spike Protein S1 Subunit and Its Coding DNA from Recombinant Middle East Respiratory Syndrome Virus

doi:10.13560/j.cnki.biotech.bull.1985.2018-0273

Abstract

Abstract: Gene fragment of S1 subunit of spike protein was artificially synthesized. Then recombinant expression plasmid Peak13CD5LTEVhIgGFc containing MERS（Middle East respiratory syndrome）coronavirus spike protein（MERS-Co V S1）was constructed and transfected into HEK293E cells,and a positive monoclonal cell line of that may stably express recombinant fusion protein was selected. The recombinant fusion protein S1-Fc with a molecular weight of 122 kD was obtained after expression and purification,and confirmed as the target protein by Western blotting. Mice were inoculated with the recombinant plasmid Peak13CD5LS1TEVhIgGFc,the recombinant S1-Fc protein,and the combination of the recombinant plasmid and S1-Fc protein,respectively. The purified recombinant S1-Fc was used as the antigen,and the titer and specificity of the antiserum of the test mice were detected by Western blotting and ELISA. The results showed that specific S1 antiserums were all present in the mice immunized with the three different inoculations. Among them,the mice immunized with the recombinant S1-Fc protein and the combination of recombinant S1-Fc protein and Peak13CD5LS1TEVhIgGFc produced high titers of antiserums against S1 after the first time of the immunization,while the mice immunized only by recombinant plasmid produced antiserum after 3 times immunizations. The antiserums prepared by immunization with the recombinant S1-Fc and the combination of recombinant S1-Fc and recombinant plasmid had the consistent antiserum titer,higher than those in the mice immunized only with the recombinant plasmid,suggesting that the inoculation of recombinant S1-Fc protein in short term was fast and efficient.

Key words: Middle East respiratory syndrome coronavirus, recombinant plasmid, recombinant S1-Fc, inoculation, serum titer

ZHANG Qian-qian, LIU Kang-tai, HAN Zong-sheng, LI Rong-gui. Antiserum Levels in Mice Inoculated with Spike Protein S1 Subunit and Its Coding DNA from Recombinant Middle East Respiratory Syndrome Virus[J]. Biotechnology Bulletin, 2018, 34(10): 150-156.

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