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
2010年
1期
30-32
,共3页
陈斯勇%郭敏卓%邱丰%费永亮%伊瑶%郭瑜%贾志远%余陶%毕胜利
陳斯勇%郭敏卓%邱豐%費永亮%伊瑤%郭瑜%賈誌遠%餘陶%畢勝利
진사용%곽민탁%구봉%비영량%이요%곽유%가지원%여도%필성리
肝炎病毒,乙型%表位%病毒粒子
肝炎病毒,乙型%錶位%病毒粒子
간염병독,을형%표위%병독입자
Hepatitis B virus%Epitopes%Virion
目的 构建一个以乙肝核心抗原为骨架的病毒样颗粒,乙肝表面抗原"a"决定簇呈现在该病毒样颗粒的表面.方法 乙肝adr血清型的HBsAg蛋白"a"抗原决定簇(aa139～148,CSKPTDGNCT)插入到HBcAg的第78位天冬氨酸和第79位脯氨酸之间,密码子优化之后基因合成,酶切后连接到表达载体上,在大肠埃希菌中进行表达,纯化后对蛋白进行电镜观察、ELISA检测抗原性、免疫兔后检测免疫原性.结果 构建得到预期的原核表达质粒,并在大肠埃希菌中表达,纯化后电镜观察为病毒样颗粒,ELISA检测证实具有良好的抗原性和免疫原性.结论 成功获得预期的带乙肝表面抗原"a"决定簇的病毒样颗粒,为其应用奠定基础.
目的 構建一箇以乙肝覈心抗原為骨架的病毒樣顆粒,乙肝錶麵抗原"a"決定簇呈現在該病毒樣顆粒的錶麵.方法 乙肝adr血清型的HBsAg蛋白"a"抗原決定簇(aa139～148,CSKPTDGNCT)插入到HBcAg的第78位天鼕氨痠和第79位脯氨痠之間,密碼子優化之後基因閤成,酶切後連接到錶達載體上,在大腸埃希菌中進行錶達,純化後對蛋白進行電鏡觀察、ELISA檢測抗原性、免疫兔後檢測免疫原性.結果 構建得到預期的原覈錶達質粒,併在大腸埃希菌中錶達,純化後電鏡觀察為病毒樣顆粒,ELISA檢測證實具有良好的抗原性和免疫原性.結論 成功穫得預期的帶乙肝錶麵抗原"a"決定簇的病毒樣顆粒,為其應用奠定基礎.
목적 구건일개이을간핵심항원위골가적병독양과립,을간표면항원"a"결정족정현재해병독양과립적표면.방법 을간adr혈청형적HBsAg단백"a"항원결정족(aa139～148,CSKPTDGNCT)삽입도HBcAg적제78위천동안산화제79위포안산지간,밀마자우화지후기인합성,매절후련접도표체재체상,재대장애희균중진행표체,순화후대단백진행전경관찰、ELISA검측항원성、면역토후검측면역원성.결과 구건득도예기적원핵표체질립,병재대장애희균중표체,순화후전경관찰위병독양과립,ELISA검측증실구유량호적항원성화면역원성.결론 성공획득예기적대을간표면항원"a"결정족적병독양과립,위기응용전정기출.
Objective To construct virus-like particles of hepatitis B core antigen, with HBsAg "a" epitope exposed on the surface. Methods Hepatitis B surface antigen "a" epitope were inserted into the Hepatitis B core antigen, between the 78th (Asp) and the 79th (Pro) amino acids. The gene was synthesized after the codon optimized, then it was ligated to the express vector after been enzyme digest. The virus-like particles were observed by electron microscope and detected by ELISA after been expressed and purified. Immune the rabbits by the VLPs, then detect the antibody. Result The virus-like particles were confirmed by electron microscope. Its antigenicity and immunogenicity were identified by ELISA. Conclusion The prokaryotic express plasmid with the fusion gene has been constructed successfully. The virus-like particles have been expressed, purified and identified, which lays the foundation for its application in the further.
