重组日本血吸虫14-3-3蛋白的纯化和免疫特性分析

The purification and the immuno properties analysis of rSj 14-3-3 protein

目的 :纯化日本血吸虫信号蛋白质 14 3 3编码基因的原核表达产物 ,分析纯化产物的免疫学特性。方法 :SDS PAGE鉴定重组日本血吸虫 14 3 3(rSj14 3 3)蛋白包涵体 ,超声破碎细胞收集包涵体 ,尿素溶解包涵体 ,NiSO4平衡层析柱 ,亲和层析纯化rSj14 3 3,Western blot鉴定纯化产物的免疫学特性。结果 :rSj14 3 3是以包涵体形式在大肠埃希菌中表达 ,通过亲和层析在 32 .5kDa附近得到单一蛋白条带 ,Western blot证实纯化产物为rSj 14 3 3,且具有与天然Sj 14 3 3相同的抗原表位。结论 :成功纯化了rSj14 3 3蛋白 ,为研究 14 3 3在血吸虫信号转导中的作用奠定了基础。

Objective:To purify the prokaryotic expression products of Schistosoma japonicum signal transduction protein 14 3 3 gene and identify the immunologic properties of the purified protein.Methods:The rSj 14 3 3 inclusion bodies were identified by SDS PAGE and collected after the E.coli cells were sonicated.To dissolve the inclusion bodies,urea was used as denaturant.The column was charged and equilibrated with NiSO 4.The rSj 14 3 3 was purified by affinity chromatography and its immunogenic properties were identified by Western blotting assay.Results:The rSj 14 3 3 was expressed in E.coli as inclusion bodies.After affinity chromatography,we obtained a single protein band at about 32.5 kDa.The result of Western blot indicated that the purified protein was rSj 14 3 3 which has the same epitopes as natural Sj 14 3 3.Conclusions:The purification of rSj 14 3 3 was succeeded and it provided a platform for the study of Schistosoma signal transduction.