J亚群禽白血病病毒SU基因与兔IgG Fc基因在Sf9细胞中的融合表达  

Fusion expression of subgroup J avian leukosis virus SU domain and Fc of rabbit IgG in Sf9 cells

作　　者：孙薇薇 秦爱建 梅梅 钱琨 金文杰 范忠军 杭柏林 尹丽萍 邵红霞 

机构地区：[1]扬州大学江苏省动物预防医学重点实验室/教育部禽类预防医学重点实验室,江苏扬州225009

出　　处：《畜牧与兽医》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

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.

关 键 词：J亚群禽白血病病毒 su基因 兔IGG FC 融合表达 

分 类 号：S852.65[农业科学—基础兽医学]