[日本血吸虫重组22.6 kDa融合蛋白的表达与鉴定]
[Expression and identification of recombinant 22.6 kDa fusion protein of Schistosoma japonicum].
作者信息
Su C, Ma L, Wu H, Shen L, Chen S, Zhang Z, Wu G
机构信息
Department of Parasitology, Nanjing Medical University, Nanjing 210029.
出版信息
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 1999;17(4):205-8.
AIM
To obtain a large amount of purified 22.6 kDa antigen of Schistosoma japonicum (Sj22.6) in large quantity.
METHODS
The sequence of the gene fragment encoding Sj22.6 was reformed by PCR and subcloned into plasmid vector pGEX-1 lambda T that coded for the 26 kDa GST antigen of Schistosoma japonicum (Sj26 GST). The recombinant plasmid was transformed into E. coli TG2 and then the positive recombinant clone was expressed by induction with IPTG.
RESULTS
The recombinant Sj22.6/Sj26 GST fusion protein was expressed in 5.1% of total bacterial protein and was easy to be purified with glutathione sepharose 4B. Moreover, the purified recombinant Sj22.6 antigen could be cut off easily from the fusion protein with thrombin and had high immunogenicity.
CONCLUSION
The purified recombinant Sj22.6 protein and Sj22.6/Sj26 GST fusion protein had the same immunological activity as the native Sj22.6 kDa protein.
目的
大量获取纯化的日本血吸虫22.6 kDa抗原（Sj22.6）。
方法
通过PCR对编码Sj22.6的基因片段序列进行改造，并亚克隆至编码日本血吸虫26 kDa谷胱甘肽S-转移酶抗原（Sj26 GST）的质粒载体pGEX-1λT中。将重组质粒转化至大肠杆菌TG2，然后用异丙基-β-D-硫代半乳糖苷（IPTG）诱导阳性重组克隆表达。
结果
重组Sj22.6/Sj26 GST融合蛋白表达量占细菌总蛋白的5.1%，且易于用谷胱甘肽琼脂糖4B进行纯化。此外，纯化的重组Sj22.6抗原可用凝血酶从融合蛋白中轻松切割下来，且具有高免疫原性。
结论
纯化的重组Sj22.6蛋白和Sj22.6/Sj26 GST融合蛋白与天然的22.6 kDa Sj22.6蛋白具有相同的免疫活性。
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