YggG截短蛋白的表达及其抗体的制备  

The Expression of truncated YggG protein and preparation of its polyclonal antibody

作　　者：黄勇[1] 陈苏民[1] 陈南春[1] 李洁[1] 张斌[1] 张晓楠[1] 

机构地区：[1]第四军医大学生物化学与分子生物学教研室,陕西西安710032

出　　处：《细胞与分子免疫学杂志》2004年第2期171-173,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目 (No .39870 380 )

摘　　要：目的 :在大肠杆菌中表达截短的YggG蛋白 ,并制备兔抗YggG截短体抗体。方法 :从含有大肠杆菌yggg基因全长DNA的质粒中 ,用PCR扩增截短的yggg基因序列 ,克隆入非融合表达载体pDH2中 ,构建YggG截短体的高表达工程菌 ,并温敏诱导表达非毒性的YggG蛋白 ,薄层扫描检测目的蛋白含量。免疫家兔制备兔抗YggG突变体抗血清 ,并以Western blot检测抗体效价、鉴定抗体特异性。结果 :大肠杆菌中表达的YggG截短突变体蛋白占菌体总蛋白的 3 1.7%。抗血清效价约 1∶2 0 0 0 ,Westernblot分析显示抗血清具有较高的特异性。结论 :成功地获得了YggG截短体蛋白 ,并制备了兔抗YggG突变体抗血清。AIM: To express truncated YggG protein(TYP) in Escherichia coli and to prepare anti-TYPE antibody. METHODS: Truncated yggg gen e was amplified by PCR from the plasmid containing full length yggg gene and was cloned into the expression vector pDH2. The expression of TYP was achieved by thermal induction. Antiserum was prepared by immunizing rabbit with TYP, T he titer and specificity of polyclonal antibody were detected by Western blot. RESULTS: Thin-layer scan analysis showed that TYP accounted fo r about 37.1% of total bacterial protein .The antiserum was about 1∶ 2 000 in titier and highly specific. Full-lergth rggG protein coul d also be recognized by the antiserum. CONCLUSION: The T YP was highly expressed, and specific anti-TYP antiserum was prepared success fully.

关 键 词：YggG蛋白 截短突变体 多克隆抗体 

分 类 号：Q754[生物学—分子生物学]