摘要
用PCR扩增方法将ALV Jenv基因不同片段进行了克隆 ,并构建了env基因片段GST融合蛋白载体。用Westernblot实验证明 ,大肠杆菌表达的不同env基因片段的GST融合蛋白能与相应的单克隆抗体产生特异性反应性 ,单克隆抗体JE9和G2识别的抗原位点位于gp85的氨基酸 6 5～ 1 5 5区域 ,而I45识别的抗原表位位于env基因的另一区域 (1 5 6～ 2 3 3位氨基酸 )。ALV J氨基酸多肽而非糖基化位点决定ALV
Envelope glycoprotein of avian leukosis virus Subgroup J (ALV-J) determines the host range of virus infection and cross-neutralization patterns.The truncated envelope genes of avian lecukosis virus subgroup J (ALV-J) were amplified by PCR and cloned them into pGEX-5X-3 vector for expressing envGST-fusion protein.Western blot analysis results showed that the products of truncated env gene expressed in Escherichia coli could reacted with G2,JE9 and I45 monoclonal antibodies (Mabs) specific to envelope protein of ALV-J.Using different Mabs to map the epitopes in the expressed truncated gp85 GST fusion protein,the results showed that Mab G2 and JE9 antibodies recognizing epitope in gp85 was localized between amino acid 65～155.Mab I45 reacted with the epitope at the location of amino acid 156～233.It indicated that the specificity of subgroup J virus is determined by the gp85 peptide since GST-gp85 protein expressed in Escherichia coli is not glycosylated.
出处
《微生物学报》
CAS
CSCD
北大核心
2002年第1期99-104,共6页
Acta Microbiologica Sinica
基金
江苏省教育厅自然基金重点项目 (OOKJB2 30 0 0 2 )
教育部基金
关键词
禽白血病病毒
J亚群
ENV基因
表达
抗原分析
Avian leukosis virus, subgroup J, Antigen analysis, Envelope gene, Envelope protein
