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采用PCR方法特异性扩增恶性疟原虫云南株(PFD-3/YN)丝氨酸重复抗原基因片段,经基因序列测定后克隆于pWR450-1融合蛋白表达载体,并转化大肠杆菌TG1。在不同菌体浓度及不同剂量IPTG诱导下检测SERA融合蛋白的表达。采用dot-ELISA及Western-blot分析对表达产物进行鉴定,结果显示SERA基因与pWR450-1重组后在大肠杆菌TG1中表达一72kDa的融合蛋白,当工程菌OD590值为0.8～1.2时,加入终浓度1mmol/LIPTG进行诱导,表达量较高。dot-ELISA和Westernblot分析表明SERA基因表达产物能被抗SERA单克隆抗体所识别。
The serine repeat antigen gene fragment of P. falciparum strain (PFD-3 / YN) was amplified by PCR and cloned into pWR450-1 fusion protein expression vector and transformed into E.coli TG1. The expression of SERA fusion protein was detected under different bacterial concentrations and different doses of IPTG. The results of dot-ELISA and Western-blot analysis showed that SERA gene was expressed in Escherichia coli TG1 after recombinant with pWR450-1, and a fusion protein of 72kDa was expressed. When OD590 of engineering strain was 0.8-1.2, The concentration of 1mmol / LIPTG induction, higher expression. Dot-ELISA and Western blot analysis showed that SERA gene expression product was recognized by anti-SERA monoclonal antibody.