Zhang Hui, Zhao Li, Ren Jiao, Gao Jian, Bian Tao, Fan Jiang-tao, Ruan Li, Chen Xin-qiu, Tian Hou-wen
Department of Gynecology, The Oncology Hospital of Guangxi Medical University, Nanning, 530021 China.
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2007 Jun;21(2):156-8.
To construct the Escherichia coli (E. coli) prokaryotic expression system pET9aHPV11L2E7, purify the fusion protein L2E7 and study the immunnogenicity of the protein.
The HPV11 L2, E7 coding region was amplified from condyloma acuminata tissue specimen by PCR. The recombinant plazmid pET9aHPV11L2E7 was established and sequenced. Fusion protein L2E7 (553 amino acids) was expressed in host strain BL21 (DE3plus) by IPTG inducing and identified by using SDS-PAGE and Western blotting. Then L2E7 protein purified with CM column was inoculated to Balb/c mice and its cell-mediated and humoral immunnogenicity was assessed by IFN-gamma enzyme-linked immunospot (ELISPOT) and enzyme-linked immunosorbent assay (ELISA).
The E. coli prokaryotic expression system pET9aHPV11L2E7 was established and the purified fusion protein L2E7 was obtained successfully. The mice in vivo experiment indicated that the purified protein L2E7 could induce HPV11E7 specific cell-mediated immune responses and high level HPV L2E7 antibody was detected in serum.
The purified fusion protein L2E7 could induce specific cell-mediated and humoral immune responses. It can be used as a candidate of genital wart immune therapeutic vaccine.
构建大肠杆菌（E. coli）原核表达系统pET9aHPV11L2E7，纯化融合蛋白L2E7并研究其免疫原性。
通过PCR从尖锐湿疣组织标本中扩增HPV11 L2、E7编码区。构建重组质粒pET9aHPV11L2E7并测序。融合蛋白L2E7（553个氨基酸）经IPTG诱导在宿主菌BL21（DE3plus）中表达，采用SDS-PAGE和Western印迹法进行鉴定。将经CM柱纯化的L2E7蛋白接种Balb/c小鼠，通过γ干扰素酶联免疫斑点试验（ELISPOT）和酶联免疫吸附试验（ELISA）评估其细胞介导免疫和体液免疫原性。
成功构建大肠杆菌原核表达系统pET9aHPV11L2E7，获得纯化的融合蛋白L2E7。小鼠体内实验表明，纯化的蛋白L2E7可诱导HPV11E7特异性细胞介导免疫反应，血清中检测到高水平的HPV L2E7抗体。
纯化的融合蛋白L2E7可诱导特异性细胞介导免疫和体液免疫反应。可作为尖锐湿疣免疫治疗疫苗的候选物。
