赤羽病毒G1基因的真核表达及抗原性检测  

Eukaryotic expression of Akabane virus G1 gene

作　　者：徐树兰[1] 童铁钢[1] 白宇[1] 王群[1] 张维军[1] 孙庆歌[1] 吴东来[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所国家兽医生物技术重点实验室人畜共患病室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2008年第6期440-444,共5页Chinese Journal of Preventive Veterinary Medicine

摘　　要：通过RT-PCR获得赤羽病毒(AKAV)OBE-1株的G1基因,克隆至pMD18-T载体。经测序鉴定正确后,将该片段作为目的基因亚克隆至Bac-to-Bac杆状病毒表达系统中的pFastBacHTA转移载体质粒中,利用Tn7转座子与穿梭载体BacmidDNA同源重组,获得了重组杆状病毒DNA,用CELLFECTIN介导其转染Sf9昆虫细胞,获得重组杆状病毒BAC-G1P1。SDS-PAGE结果表明,感染BAC-G1P1后的Sf9昆虫细胞表达可溶形式的120ku目的蛋白,其大小与预期相符。Western blot和间接免疫荧光试验显示表达产物具有良好的免疫学活性。The G1 gene of Akabane virus OBE-1 strain was amplified from the AKAV genome by RT-PCR. The PCR product was cloned into pFastBacHT A donor plasmid of Bac-to-Bac baculovirus expression system. Recombinant DNA containing the GI gene was obtained through homologous recombination of the donor plasmid with Bacmid DNA at the site of TnT. Recombinant virus BAC-GIPI was generated by transfecting recombinant DNA into Sf9 cells. SDS-PAGE analysis indicated that the GI protein expressed in Sf9 cells was approximately 120 ku in size. Western blot and indirect immunofluorescence assay （IFA） showed that the recombinant GI protein had good antigenicity.

关 键 词：赤羽病毒 G1基因 表达 

分 类 号：S852.659.5[农业科学—基础兽医学]