外源重组蛋白制备鹅免疫球蛋白轻链特异性多克隆抗体

Preparation of polyclonal antibody specific for light chain of goose im munoglobulin using recombinant protein

采用RT-PCR方法扩增鹅免疫球蛋白轻链恒定区编码序列(GoIgCL),构建原核表达载体pET30a-IgCL,在RosettaTM(DE3)pLysS宿主菌中表达鹅免疫球蛋白轻链恒定区重组蛋白rGoCL,以纯化后rGoCL作为免疫原制备兔抗GoIgL多克隆抗体,对多抗进行鉴定分析。结果表明,rGoCL在大肠杆菌中获得可溶性表达,可代替天然分离的轻链作为免疫原制备轻链特异性抗体,多抗效价为1 204 800。研究为鹅免疫球蛋白的蛋白结构、功能分析以及鹅免疫诊断试剂研发奠定基础。

In this study,the constant region of goose immunoglobulin light chain（GoIgCL） was amplified by RT-PCR.The recombinant plasmid pET30a-IgCL was constructed to express the recombinant protein rGoCL of the constant region of goose immunoglobulin light chain in RosettaTM（DE3）pLysS.The polyclonal antibody（PAb） against GoIgL was prepared with purified rGoCL,the titer and reactivity of PAb were identified.The results showed that rGoCL was expressed in E.coli as the soluble form,which could replace the natural light chain separated as the immnuogen in preparing specific antibody against GoIgL,and the titer of PAb obtained here was 1： 204800.It might facilitate further studies on the protein structure and function of goose immunoglobulin,as well as the development of the immunodiagnostic regent for goose disease.