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HJC基因是由2个Bt基因(cry1Ab和vip3)经过人工融合而成,具有更广谱的杀虫活性,可延缓害虫产生交互抗性的时间。将已构建好的携带HJC基因的重组质粒pET28a-HJC转化到大肠杆菌BL21中诱导表达。该HJC融合蛋白主要以包涵体形式存在,变性条件下使用镍亲和层析柱对其进行纯化,并经尿素梯度透析复性后,进行免疫反应活性及美国白蛾杀虫活性测定。Western blot结果显示,该原核表达蛋白与转HJC基因水稻中的HJC蛋白有相同的免疫反应性,对美国白蛾也有一定的杀虫活性,可以替代植物外源蛋白进行转HJC基因产品的食用安全性评价。
The HJC gene is synthesized by two Bt genes (cry1Ab and vip3) and has a broader spectrum of insecticidal activities, which can delay the time for the pests to develop cross-resistance. The constructed recombinant plasmid carrying HJC gene pET28a-HJC was transformed into E. coli BL21 to induce expression. The HJC fusion protein mainly exists in the form of inclusion body. The HJC fusion protein was purified by nickel affinity chromatography under denaturing conditions, and the immunoreactivity and the insecticidal activity of the American white moth were determined by urea gradient dialysis. The result of Western blot showed that the prokaryotic expression protein had the same immunoreactivity with HJC protein in HJC transgenic rice and some insecticidal activity to H. litura, which could substitute the foreign protein of plant for food safety of HJC gene product Sexual evaluation.