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【目的】构建日本血吸虫Mr=26×103谷胱甘肽S-转移酶(GST)基因的植物表达载体,为研制血吸虫病口服疫苗做前期准备。【方法】采用PCR技术,扩增目的基因GST,与植物表达载体pBI121连结,构建重组表达载体pBI121-GST。【结果】通过PCR检测和双酶切鉴定以及重组质粒序列测定,结果表明,该目的基因片段已被整合到植物表达载体pBI121中,通过电激转化,将质粒转入农杆菌菌株LBA4404、EHA105中。【结论】本实验成功地构建了日本血吸虫Mr=26×103谷胱甘肽S-转移酶GST基因的植物表达载体。
【Objective】 To construct the plant expression vector of Mr = 26 × 103 glutathione S-transferase (GST) gene of Schistosoma japonicum for the preparation of oral vaccine for schistosomiasis. 【Method】 The target gene GST was amplified by PCR and ligated with the plant expression vector pBI121 to construct the recombinant expression vector pBI121-GST. 【Result】 The results showed that the target gene fragment was integrated into the plant expression vector pBI121 by PCR assay, double enzyme digestion and sequencing of the recombinant plasmid. The plasmid was transformed into Agrobacterium strains LBA4404 and EHA105 by electroporation . 【Conclusion】 The experiment successfully constructed the plant expression vector of GST gene of Mr = 26 × 103 glutathione S-transferase of Schistosoma japonicum.