摘要
目的 本实验选择的真核表达载体 pJ16 ,是以痘苗病毒复制非必需区血凝素 (HA)基因为侧翼 ,以牛痘病毒包涵体 (ATI)启动子和串联的 p7 5突变型早期启动子为基本构件的痘苗病毒表达载体。插入编码AIDS病毒 (HIV -1)外膜蛋白env与编码干扰素IFNα - 2b基因。方法 经与野生型痘苗病毒在细胞内同源重组 ,挑选HA-斑。获得重组痘苗病毒vJ16env/IFNα - 2b。经免疫荧光、Dotblot、SDS -PAGE、Westernblot和免疫小鼠后血清抗体IgGOD值检测。结果 vJ16en/IFNα - 2b能表达Env -IFNα - 2b融合蛋白 ,分子量约 10 0kDa。免疫小鼠实验 ,血清抗体IgGOD值组间均数比较 (P <0 0 0 1) ,含IFNα- 2b与vJ6env组比较 (P >0 0 5 ) ,但有增高趋势。结论 重组病毒表达的Env -IFN融合蛋白具有良好的反应原性和免疫原性。
ObjectivepJ16 was used expression vector,Env gene encoding type Ⅰ envelope protein of AIDS virus(HIV) and encoding interferon α-2b gene of human were inserted on the downstream of the hybid promotor,where HA gene of the vaccinia virus unnecessary areas was the flank and there were 16 p7.5 promtors of mutant-type vaccinia virus.MethodsA-type inclusion later promoter of vaccinia virus and vJ2env-IFNα-2b was acquired by homologous recombination with the natural vaccinia virus an selection of the HA - plaque as substrate.ResultsThe analysis of IFA.dot blot,SDS-PAGE,Western blot and Dot-ELISA.That the recombinant viruses could express Env/IFNα-2b,whose molecular weight was 100kDa.In the mice immunization expreiment,the OD value of the serum antibodies to HIV-1( P <0.001) in groups.There was difference between contain IFN and vJ16env.ConclusionThe expressed env/IFNα-2b proteins both had good reactinogenicity and immunogenicity.
出处
《中国公共卫生》
CAS
CSCD
北大核心
2002年第12期1441-1443,共3页
Chinese Journal of Public Health
