摘要
为获得J亚群禽白血病病毒(ALV-J)SU和兔IgG Fc的融合蛋白,采用PCR方法扩增出SUJ-IgG Fc基因,并克隆至pFastBac1质粒,构建转移载体pFastBac1-SUJ-IgG Fc;再将其转化DH10BacTM感受态细胞,获得重组杆状病毒穿梭质粒rBacmid-SUJ-IgG Fc;最后转染Sf9细胞,获得重组病毒rBac-SUJ-IgG Fc。免疫荧光试验结果显示,重组杆状病毒表达的融合蛋白可被ALV-J单抗JE9以及羊抗兔IgG所识别。Western blot结果显示:表达的融合蛋白与ALV-J单抗JE9以及羊抗兔IgG都有很好的反应性,其分子量大小约为95 ku。该融合蛋白的表达为鸡细胞表面ALV-J受体的研究提供了有力工具。
To obtain the protein of subgroup J avian leukosis virus （ALV-J） SU fused to rabbit IgG Fc, the ALV-J SU and rabbit IgG Fc genes were amplified by PCR and cloned into pFastBacl to construct the recombinant plasmid pFastBacl-SUJ-IgG Fc. Then, pFastBacl-SUJ- IgG Fc was transformed into DH10BacTM competent cells to construct the recombinant shuttle plasmid. The shuttle plasmid was transfected to Sf9 cells to get recombinant baculovirus. Immunofluorescence assay showed that the fusion protein could be recognized by JE9 monoclonal antibody and antibodies to rabbit IgG. Western-blot results showed that the molecular weight of fusion protein was about 95ku. The expression of fusion protein provides a powerful tool to study ALV-J receptor on the surface of chicken cells.
出处
《畜牧与兽医》
北大核心
2013年第3期1-5,共5页
Animal Husbandry & Veterinary Medicine
基金
联合国家基金重点项目(U0831002)
国家基金青年项目(31201881)
教育部创新团队项目(IRT0978)
江苏省优势学科项目
