J亚群禽白血病病毒膜表面糖蛋白基因gp85的克隆与表达

Cloning and expression of env-gp85 of ALV-J IMC_(10200) strain

以pGEM IMC2 .2 重组质粒为模板扩增禽白血病病毒内蒙株IMC10 2 0 0 gp85基因 ,构建了转移载体pFastBacl IMCgp85 ,并利用Bac to Bac表达系统获得了重组杆状病毒rBac IMCgp85。转染Sf 9细胞后的表达研究表明 ,重组杆状病毒可高效表达IMC10 2 0 0 的gp85基因 ,表达产物具有病毒的抗原特异性 ,与ALV J单抗JE 9有强的间接免疫荧光反应。蛋白质印迹法分析表明 ,表达产物的分子质量约 5 4ku。

This report described the cloning and expression of envelope gene gp85 of IMC 10200 strain of avian leukosis virus subgroup J (ALV J). The sequence encoding the gp85 domain of ALV J IMC 10200 strain was amplified from pGEM IMC 2.2 vector, and cloned into vector pFast Bacl. By the Bac to Bac baculovirus expression system, rBac IMC gp85 was obtained. Immunofluorescence assay with JE 9 monoclonal antibody against ALV J showed that the recombinant gp85 gene product was expressed in Sf9 cells infected with rBac IMC gp85. The molecular weight of expressed protein was about 54 ku by western blotting.