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应用聚合酶链反应技术 ,从日本血吸虫 (大陆株 ) 2 3kD分子基因中扩增出其大亲水肽段的DNA序列 ,将此片段插入到表达载体 pGEX 5X 1中进行表达 ,经IPTG诱导产生分子量约 33 5kD的融合蛋白 (GST HD)。用FactorXa对融合蛋白进行切割获得纯化的大亲水肽段 ,Westernblotting分析表明 ,融合蛋白及大亲水肽段均能为血吸虫病人血清识别。
Polymerase chain reaction (PCR) was used to amplify the DNA sequence of the large hydrophilic peptide fragment from 23kD gene of Schistosoma japonicum (Chinese strain). The fragment was inserted into the expression vector pGEX 5X 1 and induced by IPTG A molecular weight of about 33 5kD fusion protein (GST HD). The fusion protein was cleaved with FactorXa to obtain the purified large hydrophilic peptides. Western blotting analysis showed that both the fusion protein and the large hydrophilic peptides could recognize the serum of schistosomiasis patients.