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作 者:Fethia Ben Yebdri Abderrahmane AAZAZ 叶凯[1] 马辉文[1] 童立恒
机构地区:[1]武汉大学药学院,武汉430072 [2]武汉生物技术研究开发中心,武汉430030
出 处:《生物工程学报》2004年第5期683-688,共6页Chinese Journal of Biotechnology
摘 要:研究了庚型肝炎病毒E2 (HGVE2 )基因片段作为DNA疫苗的可行性。将来自于质粒pThioHis E2编码HGVE2的基因片段 (5 5 9bp)亚克隆到质粒pCMV S中 ,使之和HBsAg基因位于同一阅读框 ,形成重组质粒pCMV S E2。用纯化的质粒pCMV S E2DNA注射到昆明小鼠后腿四头肌中来免疫小鼠 ,同时用pCMV S作为对照。间隔 14天再加强一次免疫。在加强免疫后的第 8天眼眶取血。用E2 GST融合蛋白作为固定化抗原 ,通过ELISA检测受试小鼠的体液免疫应答。结果表明 ,用质粒pCMV SIn order to study the feasibility of E2 gene fragment of hepatitis virus G(HGV) as a component of DNA vaccine against the hepatitis virus G infection, a 559bp DNA fragment encoding HGV E2 was cloned into plasmid pCMV S from pThioHis E2 in the same reading frame with HBsAg gene to form a recombinant plasmid named pCMV S E2 BALB/c mice of Kunming strain were immunized with purified plasmid DNA of pCMV S E2 by intra muscularly inoculation. The immunizations were boosted twice at an interval of 14 days. The whole blood was collected from mice orbit on the day 8 after the last boost. Mice sera were screened by ELISA to determine the humoral immune response using E2 GST fusion protein as the immobilized antigen and the sera from mice immunized with pCMV S as control. The result indicated that the immunization with plasmid DNA of pCMV S E2 could induce quite strong humoral immune response.
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