生物技术通讯
生物技術通訊
생물기술통신
LETTERS IN BIOTECHNOLOGY
2015年
2期
194-198
,共5页
王平兴%任皎%赵莉%邓瑶%阳静%李善琴%阮力%谭文杰%田厚文
王平興%任皎%趙莉%鄧瑤%暘靜%李善琴%阮力%譚文傑%田厚文
왕평흥%임교%조리%산요%양정%리선금%원력%담문걸%전후문
中东呼吸综合征冠状病毒%病毒载体%疫苗%棘突蛋白
中東呼吸綜閤徵冠狀病毒%病毒載體%疫苗%棘突蛋白
중동호흡종합정관상병독%병독재체%역묘%극돌단백
Middle East respiratory syndrome coronavirus%virus vector%vaccines%spike protein
目的:以痘苗病毒天坛株为载体,构建表达中东呼吸综合征冠状病毒(MERS-CoV)S蛋白的重组病毒疫苗,并进行免疫效果评价。方法:通过PCR扩增获得去除跨膜区的S蛋白基因片段SQ,并构建重组痘苗病毒载体质粒pJSC11LacZ7.5SQ,将重组质粒与痘苗病毒同源重组并单斑纯化获得重组病毒毒株RVVMERS-SQ,重组病毒免疫小鼠后用酶联免疫吸附实验(ELISA)和假病毒微量中和实验检测其诱发的S蛋白体液免疫反应水平。结果:构建了表达MERS-CoV去跨膜区S蛋白的重组病毒RVVMERS-SQ,其免疫小鼠后诱发了强的S蛋白体液免疫反应,血清IgG抗体滴度为1∶3200,中和抗体滴度达到1∶1000。结论:重组痘苗病毒RVVMERS-SQ可在BALB/c小鼠体内诱发强的免疫反应,为MERS-CoV疫苗的研发提供了实验基础。
目的:以痘苗病毒天罈株為載體,構建錶達中東呼吸綜閤徵冠狀病毒(MERS-CoV)S蛋白的重組病毒疫苗,併進行免疫效果評價。方法:通過PCR擴增穫得去除跨膜區的S蛋白基因片段SQ,併構建重組痘苗病毒載體質粒pJSC11LacZ7.5SQ,將重組質粒與痘苗病毒同源重組併單斑純化穫得重組病毒毒株RVVMERS-SQ,重組病毒免疫小鼠後用酶聯免疫吸附實驗(ELISA)和假病毒微量中和實驗檢測其誘髮的S蛋白體液免疫反應水平。結果:構建瞭錶達MERS-CoV去跨膜區S蛋白的重組病毒RVVMERS-SQ,其免疫小鼠後誘髮瞭彊的S蛋白體液免疫反應,血清IgG抗體滴度為1∶3200,中和抗體滴度達到1∶1000。結論:重組痘苗病毒RVVMERS-SQ可在BALB/c小鼠體內誘髮彊的免疫反應,為MERS-CoV疫苗的研髮提供瞭實驗基礎。
목적:이두묘병독천단주위재체,구건표체중동호흡종합정관상병독(MERS-CoV)S단백적중조병독역묘,병진행면역효과평개。방법:통과PCR확증획득거제과막구적S단백기인편단SQ,병구건중조두묘병독재체질립pJSC11LacZ7.5SQ,장중조질립여두묘병독동원중조병단반순화획득중조병독독주RVVMERS-SQ,중조병독면역소서후용매련면역흡부실험(ELISA)화가병독미량중화실험검측기유발적S단백체액면역반응수평。결과:구건료표체MERS-CoV거과막구S단백적중조병독RVVMERS-SQ,기면역소서후유발료강적S단백체액면역반응,혈청IgG항체적도위1∶3200,중화항체적도체도1∶1000。결론:중조두묘병독RVVMERS-SQ가재BALB/c소서체내유발강적면역반응,위MERS-CoV역묘적연발제공료실험기출。
Objective: To construct a recombinant vaccinia virus expressing middle east respiratory syndrome coronavirus(MERS-CoV) S protein and evaluate its immunogenicity. Methods: SQ fragment was amplified by PCR in which the transmembrane region of S protein was deleted, and then inserted into a vaccinia virus transfer pla?simid. The recombinant vaccinia virus expressing MERS-CoV S protein was constructed through homologous recom?binant based on Tiantan vaccinia virus strains in chicken embryo fibroblast cells. BALB/c mice were immunized with the recombinant virus, its humoral immune responses against MERS-CoV S protein were detected by enzyme-linked immunoabsorbent assay(ELISA) and pseudovirus micro-neutralization assay. Results: The recombinant vac?cinia virus(RVVMERS-SQ) expressing SQ protein was constructed. Mice vaccinated with RVVMERS-SQ produced strong humoral immune response against MERS-CoV S; and could be elicited high level of serum antibodies neu?tralizing MERS-CoV pseudovirus. Conclusion: Recombinant vaccinia virus RVVMERS-SQ could induce a robust humoral immune response against MERS-CoV S in vaccinated mice, it may serve for further study of a vaccine against MERS-CoV.