中国预防兽医学报
中國預防獸醫學報
중국예방수의학보
CHINESE JOURNAL OF PREVENTIVE VETERINARY MEDICINE
2010年
1期
56-60
,共5页
闫丽萍%华荣虹%亓文宝%周艳君%李国新%于海%姜一峰%田志军%童光志
閆麗萍%華榮虹%亓文寶%週豔君%李國新%于海%薑一峰%田誌軍%童光誌
염려평%화영홍%기문보%주염군%리국신%우해%강일봉%전지군%동광지
乙型脑炎病毒%E蛋白%抗原表位
乙型腦炎病毒%E蛋白%抗原錶位
을형뇌염병독%E단백%항원표위
Japanese encephalitis virus%envelope protein%epitope
为鉴定乙型脑炎病毒(JEV)E蛋白Ⅰ,Ⅱ结构域抗原表位,本研究设计了一系列针对E蛋白的Ⅰ、Ⅱ结构域(1 aa～296 aa)部分重叠短肽,分别进行GST融合表达,用JEV阳性血清进行western blot和ELISA反应性筛选,获得了5个线性抗原表位分别为:E1(~1FNCLGMGNRDFIEGAS~(16))、E10-4(~(77)TGEAHNEKS~(84))、E11-2(~(83)EKRADSSYVCKQ~(94))、E19(~(145)GTTTSENHGNYSAQVG~(160))和E33(~(257)SQEGGLHHALAGAIVV~(272)).除E10和E19外,其它表位均为第一次通过实验方法确定的.将这5个融合蛋白对小鼠进行免疫,获得了高滴度的针对5个融合短肽的抗血清,研究证明了这些表位具有良好的免疫原性.本研究为乙型脑炎特异性诊断试剂及表位疫苗的开发奠定了基础.
為鑒定乙型腦炎病毒(JEV)E蛋白Ⅰ,Ⅱ結構域抗原錶位,本研究設計瞭一繫列針對E蛋白的Ⅰ、Ⅱ結構域(1 aa～296 aa)部分重疊短肽,分彆進行GST融閤錶達,用JEV暘性血清進行western blot和ELISA反應性篩選,穫得瞭5箇線性抗原錶位分彆為:E1(~1FNCLGMGNRDFIEGAS~(16))、E10-4(~(77)TGEAHNEKS~(84))、E11-2(~(83)EKRADSSYVCKQ~(94))、E19(~(145)GTTTSENHGNYSAQVG~(160))和E33(~(257)SQEGGLHHALAGAIVV~(272)).除E10和E19外,其它錶位均為第一次通過實驗方法確定的.將這5箇融閤蛋白對小鼠進行免疫,穫得瞭高滴度的針對5箇融閤短肽的抗血清,研究證明瞭這些錶位具有良好的免疫原性.本研究為乙型腦炎特異性診斷試劑及錶位疫苗的開髮奠定瞭基礎.
위감정을형뇌염병독(JEV)E단백Ⅰ,Ⅱ결구역항원표위,본연구설계료일계렬침대E단백적Ⅰ、Ⅱ결구역(1 aa～296 aa)부분중첩단태,분별진행GST융합표체,용JEV양성혈청진행western blot화ELISA반응성사선,획득료5개선성항원표위분별위:E1(~1FNCLGMGNRDFIEGAS~(16))、E10-4(~(77)TGEAHNEKS~(84))、E11-2(~(83)EKRADSSYVCKQ~(94))、E19(~(145)GTTTSENHGNYSAQVG~(160))화E33(~(257)SQEGGLHHALAGAIVV~(272)).제E10화E19외,기타표위균위제일차통과실험방법학정적.장저5개융합단백대소서진행면역,획득료고적도적침대5개융합단태적항혈청,연구증명료저사표위구유량호적면역원성.본연구위을형뇌염특이성진단시제급표위역묘적개발전정료기출.
In order to map the antigenic epitopes domain Ⅰ,Ⅱ of the envelope (E) protein of Japanese encephalitis virus (JEV), a set of partially overlapping fragments spanning the E protein were fused with GST and expressed. The reactivity of the fusion proteins was detected by westem blot and ELISA. Five linear antigenic epitopes, E1 (~1FNCLGMGNRDFIEGAS~(16)), E10-4 (~(77)TGEA HNEK~(84)), E11-2 (~(83)EKRADSSYVCKQ~94)), E19 (~(145)GTTTSENHGNYSAQVG~(160)) and E33 (~(257)SQEGGLHHALAGAIVV~(272)) wue identified. Immunization of mice with the fusion proteins revealed that all five proteins could elicit short peptide specific antisera. These results provided important basis for study of the structure and function of domain Ⅰ,Ⅱ of the JEV E protein, and development of diagnostic techniques.
