免疫学杂志
免疫學雜誌
면역학잡지
IMMUNOLOGICAL JOURNAL
2001年
1期
13-16
,共4页
张应玖%金宁一%金善荣%王宏伟%沈家骢
張應玖%金寧一%金善榮%王宏偉%瀋傢驄
장응구%금저일%금선영%왕굉위%침가총
HIV-2%gp105%gp36%杆状病毒%昆虫细胞
HIV-2%gp105%gp36%桿狀病毒%昆蟲細胞
HIV-2%gp105%gp36%간상병독%곤충세포
目的 用细菌／杆状病毒（BactoBac）表达系统在昆虫细胞Sf9中表达HIV-2外膜糖蛋白gp105和跨膜糖蛋白gp36，为研制艾滋病疫苗和诊断试剂奠定基础。方法 分别将HIV-2外膜蛋白gp105和跨膜蛋白gp36全基因序列克隆到杆状病毒转座载体pFastBacHTa和pFastBacHTb中多角体启动子下游，构建成重组转座载体pFastBacHTa-gp105和pFastBacHTb-gp36，利用细菌／杆状病毒（BactoBac）表达系统筛选重组杆状病毒，并在昆虫细胞Sf9中表达HIV-2的gp105和gp36。结果 SDS-PAGE分析结果表明，gp105基因表达产物为一66000u糖蛋白，gp36基因则表达一41000u糖蛋白，与天然产物一致。Westernblot结果显示：两者均具有抗原特异性。结论 gp105和gp36在昆虫细胞中得到了高效表达，并均被糖基化；gp105接近天然产物，gp36与天然产物一致。
目的 用細菌／桿狀病毒（BactoBac）錶達繫統在昆蟲細胞Sf9中錶達HIV-2外膜糖蛋白gp105和跨膜糖蛋白gp36，為研製艾滋病疫苗和診斷試劑奠定基礎。方法 分彆將HIV-2外膜蛋白gp105和跨膜蛋白gp36全基因序列剋隆到桿狀病毒轉座載體pFastBacHTa和pFastBacHTb中多角體啟動子下遊，構建成重組轉座載體pFastBacHTa-gp105和pFastBacHTb-gp36，利用細菌／桿狀病毒（BactoBac）錶達繫統篩選重組桿狀病毒，併在昆蟲細胞Sf9中錶達HIV-2的gp105和gp36。結果 SDS-PAGE分析結果錶明，gp105基因錶達產物為一66000u糖蛋白，gp36基因則錶達一41000u糖蛋白，與天然產物一緻。Westernblot結果顯示：兩者均具有抗原特異性。結論 gp105和gp36在昆蟲細胞中得到瞭高效錶達，併均被糖基化；gp105接近天然產物，gp36與天然產物一緻。
목적 용세균／간상병독（BactoBac）표체계통재곤충세포Sf9중표체HIV-2외막당단백gp105화과막당단백gp36，위연제애자병역묘화진단시제전정기출。방법 분별장HIV-2외막단백gp105화과막단백gp36전기인서렬극륭도간상병독전좌재체pFastBacHTa화pFastBacHTb중다각체계동자하유，구건성중조전좌재체pFastBacHTa-gp105화pFastBacHTb-gp36，이용세균／간상병독（BactoBac）표체계통사선중조간상병독，병재곤충세포Sf9중표체HIV-2적gp105화gp36。결과 SDS-PAGE분석결과표명，gp105기인표체산물위일66000u당단백，gp36기인칙표체일41000u당단백，여천연산물일치。Westernblot결과현시：량자균구유항원특이성。결론 gp105화gp36재곤충세포중득도료고효표체，병균피당기화；gp105접근천연산물，gp36여천연산물일치。
Objective To develop effective vaccine and diagnostic agents ofAIDS， the external glycoprotein gp105 and transmembrane glycoprotein gp36 of human immunodeficiency virus type 2 were expressed in insect cells Spodoptera frugiperda (Sf9) using baculovirus expression system ： Bac-to-Bac system. Methods The recombinant transfer vector pFast Bac HTa-gp105 and pFast Bac HTb-gp36 were constructed by cloning HIV-2 gp105 gene and gp36 gene into the end of polyhedrin promoter of baculovirus transfer vector pFast Bac HTa and pFast Bac HTb respectively. HIV-2 external glycoprotein gp105 and transmembrane glycoprotein gp36 were expressed in insect cells Spodoptera frugiperda (Sf9) by transposing these two foreign genes to shuttle vector bacmid respectively using Bac-to-Bac system. Results The resulting products determined by SDS-PAGE showed that recombinant baculovirusescontaining gp105 gene expressed a 66 kDa glycoprotein， whereas those contai-ning gp36 gene generated a 41 000u glycoprotein， the apparent molecular weight of which is equivalent to that of natural gp36. Western blot indicated that both gp105 and gp36 showed antigenic specificity. Conclusions gp105 and gp36 has been expressed in insect cells Spodoptera frugiperda (Sf9) using baculovirus expression system.
