日本血吸虫22.6ku膜相关蛋白基因的克隆及原核表达

Molecular Cloning and Prokaryotic Expression of the Schistosoma japonicum 22.6ku Tegumental Membrane-associated Protein

为了得到重组的日本血吸虫22.6 ku膜相关蛋白,研究设计了1对引物,以日本血吸虫mRNA为模板,用RT-PCR方法克隆了576 bp的22.6 ku膜相关蛋白基因片段,测序结果表明,该片段与已公布的Sj22.6基因序列(U75510)同源性达到99%。将该基因片段插入原核表达载体pGEX-KG构建了原核表达质粒pGEX-Sj22.6,在大肠杆菌BL21 codn plus中表达获得了约48 ku的融合蛋白,经Western-blot检测,该重组蛋白具有良好的免疫原性。

In order to get the recombinant Schistosoma japonicum 22. 6 ku tegumental protein, a pair of primers was designed. The 576 bp fragment was cloned by using reverse transcirption polymerase chain reaction （RT-PCR） from the mRNA of Schistosoma japonicum. Sequence analysis revealed that this fragment shared 99% identity with the published Sj22.6 gene sequence （U75510）. The recombinant expressing plasmid pGEX-Sj22.6 was constructed and subsequently expressed in E. coli EL21. A 48 ku rSj22.6 protein was obtained. The Western-blot assay suggested that the fusion protein has good immunogenicity.