中华实验和临床病毒学杂志
中華實驗和臨床病毒學雜誌
중화실험화림상병독학잡지
CHINESE JOURNAL OF EXPERIMENTAL AND CLINICAL VIROLOGY
2013年
2期
105-108
,共4页
周玲%任皎%赵莉%冯靖%郝明强%谭文杰%阮力%曲芃芃%田厚文
週玲%任皎%趙莉%馮靖%郝明彊%譚文傑%阮力%麯芃芃%田厚文
주령%임교%조리%풍정%학명강%담문걸%원력%곡봉봉%전후문
乳头状瘤病毒,人%疫苗%表达的序列标记%中和试验
乳頭狀瘤病毒,人%疫苗%錶達的序列標記%中和試驗
유두상류병독,인%역묘%표체적서렬표기%중화시험
Papillomavirus,human%Vaccines%Expressed sequence tags%Neutralization tests
目的 高效表达HPV31和52型L2融合蛋白疫苗,并评价其免疫效果.方法 根据GenBank上公布的HPV31、52型L2蛋白11-200位氨基酸序列,设计合成两个型别L2该区域对应的优化密码子基因融合序列,并将其克隆至原核表达载体中.利用原核表达系统表达HPV31和52 L2融合蛋白,纯化目的蛋白后免疫小鼠,用血清抗体检测及HPV假病毒体外中和试验评价融合蛋白疫苗的免疫效果.结果 HPV31和52 L2融合蛋白在原核表达体系中呈高效表达,表达量约占全菌20％.融合蛋白加铝佐剂免疫小鼠后,血清中能检测到高滴度的总抗体及一定水平的中和抗体和交叉中和抗体.结论 HPV31和52 L2融合蛋白能够刺激机体产生广谱中和抗体,为高危型广谱HPVL2蛋白疫苗研发提供了实验室依据.
目的 高效錶達HPV31和52型L2融閤蛋白疫苗,併評價其免疫效果.方法 根據GenBank上公佈的HPV31、52型L2蛋白11-200位氨基痠序列,設計閤成兩箇型彆L2該區域對應的優化密碼子基因融閤序列,併將其剋隆至原覈錶達載體中.利用原覈錶達繫統錶達HPV31和52 L2融閤蛋白,純化目的蛋白後免疫小鼠,用血清抗體檢測及HPV假病毒體外中和試驗評價融閤蛋白疫苗的免疫效果.結果 HPV31和52 L2融閤蛋白在原覈錶達體繫中呈高效錶達,錶達量約佔全菌20％.融閤蛋白加鋁佐劑免疫小鼠後,血清中能檢測到高滴度的總抗體及一定水平的中和抗體和交扠中和抗體.結論 HPV31和52 L2融閤蛋白能夠刺激機體產生廣譜中和抗體,為高危型廣譜HPVL2蛋白疫苗研髮提供瞭實驗室依據.
목적 고효표체HPV31화52형L2융합단백역묘,병평개기면역효과.방법 근거GenBank상공포적HPV31、52형L2단백11-200위안기산서렬,설계합성량개형별L2해구역대응적우화밀마자기인융합서렬,병장기극륭지원핵표체재체중.이용원핵표체계통표체HPV31화52 L2융합단백,순화목적단백후면역소서,용혈청항체검측급HPV가병독체외중화시험평개융합단백역묘적면역효과.결과 HPV31화52 L2융합단백재원핵표체체계중정고효표체,표체량약점전균20％.융합단백가려좌제면역소서후,혈청중능검측도고적도적총항체급일정수평적중화항체화교차중화항체.결론 HPV31화52 L2융합단백능구자격궤체산생엄보중화항체,위고위형엄보HPVL2단백역묘연발제공료실험실의거.
Objective To express HPV31 and 52 L2 fusion protein and detect its immunogenicity.Methods According to the amino acid sequences of HPV31 and 52 L2 11-200AA published in the GenBank database,weartificially synthesized the HPV31 and 52 L2 fusion gene which was optimized according to Escherichia coli codon usage and encodes 11-200 amino acid of HPV31 and HPV52 L2,then cloned it into pET-9a vector.The HPV31 and 52 L2 fusion protein was expressed in Prokaryotic expression system and the mice were immunized with the fusion protein after purification.The immunogenicity was characterized in vaccinated mice.Results HPV31 and 52 L2 fusion protein was highly expressed in E.coli,the amount of fusion protein is nearly 20％ of the total bacterial protein.The purified fusion protein with aluminum adjuvant could induce specific high titer of IgG antibodies detected by ELISA,and also induce the neutralizing antibodies against pseudovirus of HPV31 and HPV52 and cross-neutralizing antibodies against pseudovirus of HPV45,58,16,18.Conclusion HPV31 and 52 L2 fusion protein could induce neutralizing and cross-neutralizing antibodies against HPV pseudovirus.It provides laboratory basis for development of HPV L2 protein vaccine.
