CAJ | 학술논문

为应用Bac-to-Bac杆状病毒表达系统表达小反刍兽疫病毒西藏分离株 Tibet 07的磷蛋白并对其进行鉴定，应用PCR技术扩增小反刍兽疫病毒Tibet 07株P基因片段，克隆至pFastBac/CT-TOPO载体中，构建重组穿梭质粒pFastBac-PPRV-P ，进一步构建重组杆状病毒Bacmid-PPRV-P ，转染sf 9细胞，获得重组杆状病毒。通过SDS-PAGE鉴定P蛋白的表达，利用Western blot和间接免疫荧光鉴定P蛋白的抗原性。结果表明，表达的P蛋白分子质量约54 ku；感染的sf 9细胞可与小反刍兽疫阳性血清发生特异性反应。表明应用Bac-to-Bac杆状病毒表达系统成功表达了小反刍兽疫病毒西藏分离株 Tibet 07的P蛋白，为进一步研究其生物学功能奠定了基础。
위응용Bac-to-Bac간상병독표체계통표체소반추수역병독서장분리주 Tibet 07적린단백병대기진행감정，응용PCR기술확증소반추수역병독Tibet 07주P기인편단，극륭지pFastBac/CT-TOPO재체중，구건중조천사질립pFastBac-PPRV-P ，진일보구건중조간상병독Bacmid-PPRV-P ，전염sf 9세포，획득중조간상병독。통과SDS-PAGE감정P단백적표체，이용Western blot화간접면역형광감정P단백적항원성。결과표명，표체적P단백분자질량약54 ku；감염적sf 9세포가여소반추수역양성혈청발생특이성반응。표명응용Bac-to-Bac간상병독표체계통성공표체료소반추수역병독서장분리주 Tibet 07적P단백，위진일보연구기생물학공능전정료기출。
In order to obtain purified peste des petits ruminants virus P protein ,the P gene of peste des pe-tits ruminants virus strain Tibet 07 was cloned into transfer vector pFastBac/CT-TOPO ,plasmid pFast-Bac-PPRV-P was then transformed into DHl0Bac complement cells .The recombinant bacmid DNA was i-solated and transfected into sf9 insect cells to express P protein .The expressed P protein was identified by using SDS-PAGE ,Western blot and indirect immunofluorescence assay .The results showed that the re-combinant P protein was successfully expressed in insect cells with a relative molecular of 54 ku .This work provides basis for studying its biological function .