CAJ | 학술논문

目的原核表达人乳头瘤病毒(HPV)6型L2AN360E7E6融合蛋白并对其免疫效果进行初步评价.方法用重叠PCR将HPV6b 12(1～360 bp)、E7、E6三个基因片段融合,原核表达HPV6bL2△N360E7E6融合蛋白,蛋白纯化后与Al(OH)3、CpG佐剂配伍肌内注射免疫C57BL/6小鼠,使用IFN-γ ELISPOT与ELISA分别对其细胞免疫和体液免疫效果进行评价.结果蛋白+CpG佐剂组与其他免疫组相比,针对E7与E6均有明显较强的细胞免疫反应;各免疫组均能检测到高滴度的抗L2的抗体,但各组之间无明显差异.结论利用pQE30原核表达系统成功克隆、表达和纯化了HPV6bL2△N360E7E6融合蛋白,且该蛋白与合适佐剂配伍能在C57BL/6小鼠体内诱发强的细胞免疫和体液免疫反应,为该蛋白的后期研究奠定了基础.
목적 원핵표체인유두류병독(HPV)6형L2AN360E7E6융합단백병대기면역효과진행초보평개.방법 용중첩PCR장HPV6b 12(1～360 bp)、E7、E6삼개기인편단융합,원핵표체HPV6bL2△N360E7E6융합단백,단백순화후여Al(OH)3、CpG좌제배오기내주사면역C57BL/6소서,사용IFN-γ ELISPOT여ELISA분별대기세포면역화체액면역효과진행평개.결과 단백+CpG좌제조여기타면역조상비,침대E7여E6균유명현교강적세포면역반응;각면역조균능검측도고적도적항L2적항체,단각조지간무명현차이.결론 이용pQE30원핵표체계통성공극륭、표체화순화료HPV6bL2△N360E7E6융합단백,차해단백여합괄좌제배오능재C57BL/6소서체내유발강적세포면역화체액면역반응,위해단백적후기연구전정료기출.
Objective To express HPV6bL2△N360E7E6 fusion protein in E.coli and preliminarily evaluate its immune effect.Methods Three HPV6b gene fragments,which were L2(1-360 bp),E7 and E6,were fused by overlapping PCR,then were inserted into a prokaryotic expression vector and expressed in E.coli.C57BL/6 mice were immunized with purified fusion protein plus Al(OH)3 and/or CpG adjuvants through intramuscular route,the cellular and humoral immune responses were detected by IFN-γ ELISPOT and ELISA respectively.Results Protein plus CpG adjuvant could induce the strongest cellular immune response to E7 and E6,high antibody titer against L2 could be detected in all immunized groups but there were no significant difference among these groups.Conclutions HPV6bL2△N360E7E6 gene was successfully cloned into pQE30 vector and expressed in E.coli,the fusion protein was also purified and proved that could induce strong cellular and humoral immune responses with appropriate adjuvant in C57 BL/ 6 mice and could be used for future research.