【摘 要】
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Objective: To construct the recombinant yeast expressing Pre S212o-146 and HBsAg of Hepatitis B Virus, and evaluate its immune effects of whole yeast cells.Methods: Optimized PreS2120-146 and HBsAg ge
【机 构】
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Institute of Molecular Virology and Immunology, Department of Microbiology and Immunology,Wenzhou Me
【出 处】
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中国免疫学会第九届全国免疫学学术大会
论文部分内容阅读
Objective: To construct the recombinant yeast expressing Pre S212o-146 and HBsAg of Hepatitis B Virus, and evaluate its immune effects of whole yeast cells.Methods: Optimized PreS2120-146 and HBsAg genes of HBV according to the yeast cell codon preference was synthesized and cloned into pPIC3.5K yeast expression vector.After linearized with Bgl Ⅱ restriction enzyme, pPIC3.5K/Pre S2-HBsAg recombinant plasmid was electricity transformed into GS 115 strain to construct Pre S2-HBsAg--recombinant Pichia pastoris.Identified recombinant strains were then induced by methanol.The expression of Pre S2-HBsAg was identified by SDS-PAGE, Western Blot and ELISA analysis.Heat-killed whole recombinant yeast vaccinated mice by subcutaneous injection.Specific antibodies to HBsAg were detected by ELISA, and CTL response were detected by investigating the IFN-γ expression byRT-PCR when immune spleen cells of mice were stimulated by HBsAg CTL epitope.
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