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在大肠杆菌中高效表达日本血吸虫 (Schistosomajaponicum ,Sj)Sj -Ts1融合蛋白并测定其免疫保护效果。将Sj-Ts1基因亚克隆至 pGEX - 5X - 3原核载体 ,转化入大肠杆菌ER2 5 6 6 ,并用IPTG对该重组菌进行诱导表达中。将此表达产物进行WesternBlot分析后免疫小鼠 ,免疫剂量为 10 0 μg/次 /鼠。并设蛋白佐剂对照和PBS对照。免疫三次后进行攻击感染 ,计数虫负荷及肝卵负荷。在IPTG诱导下 ,亚克隆至 pGEX - 5X - 3的Sj-Ts1基因在大肠杆菌内高效表达融合蛋白SjGST -Ts1,用此融合蛋白免疫小鼠 ,能诱导产生 2 4 16 %的减虫率和 4 8 74 %的减卵率。Sj-Ts1基因亚克隆至 pGEX -5X - 3载体后可在大肠杆菌中高效表达 ,表达产物可诱导小鼠产生一定程度的抗Sj保护性免疫力。
The Schistosoma japonicum (Sj) Sj-Ts1 fusion protein was highly expressed in Escherichia coli and its immunoprotective effect was determined. The Sj-Ts1 gene was subcloned into prokaryotic vector pGEX - 5X - 3 and transformed into E.coli ER2 566. The recombinant strain was induced to express by IPTG. The expression product was analyzed by WesternBlot after immunization of mice, the immune dose of 100 μg / time / mouse. And protein adjuvant control and PBS control. The challenge was challenged three times after immunization, and the insect load and liver egg load were counted. Under the induction of IPTG, the Sj-Ts1 gene subcloned into pGEX - 5X - 3 was highly expressed in Escherichia coli, and the fusion protein SjGST - Ts1 was highly expressed in E. coli. The immunization of mice with this fusion protein resulted in a 24.6% 4 8 74% of the egg reduction rate. The Sj-Ts1 gene was subcloned into pGEX-5X-3 vector and expressed efficiently in E. coli. The expression product of Sj-Ts1 could induce some degree of anti-Sj protective immunity in mice.