YggG截短蛋白的表达及其抗体的制备

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。用制备的该抗血清可检测到野生型全长YggG蛋白的表达

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

AIM: To express truncated YggG protein (TYP) in Escherichia coli and to prepare anti-TYPE antibody. METHODS: Truncated yggg gene 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,The titer and specificity of polyclonal antibody were detected by Western blot. RESULTS: Thin-layer scan analysis showed that TYP accounted for about 37.1% of total bacterial protein.The antiserum was about 1:2,000 in titier and highly specific. Full-length rggG protein could also be recognized by the antiserum. CONCLUSION: The TYP was highly expressed, and specific anti-TYP antiserum was prepared successfully.