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目的获得ＩＬ－２绿脓杆菌外毒素Ａ嵌合蛋白，用于相关疾病的治疗。方法用ＰＣＲ方法扩增分 离ＩＬ－２和ＰＥＡ的基因，将ＩＬ－２基因的３’端与绿脓杆菌外毒素Ａ基因的５’端连接后克隆至ｐＢＶ２２０上，转化大肠杆菌 ＤＨ５ａ。挑取菌落培养，快速提取质粒进行酶切，并在温控条件下表达。结果经鉴定，获得重组质粒ｐＢＶ／ＩＬ－２／ ＰＥＡ。温控诱导表达后ＳＤＳ－ＰＡＧＥ分析表明，在５１ ０００处有蛋白表达带。以鼠抗人ＩＬ－２单克隆抗体经免疫印迹验 证，该表达蛋白为所设计的嵌合蛋白。结论本研究建立了一种制备ＩＬ－２／ＰＥＡ嵌合蛋白的方法，该方法也可用于 表达其他蛋白。
Objective To obtain IL-2 Pseudomonas exotoxin A chimeric protein for the treatment of related diseases. Methods The gene of IL-2 and PEA was amplified by PCR. The 3 ’end of IL-2 gene was ligated to the 5’ end of toxin A gene of Pseudomonas aeruginosa and cloned into pBV220 for transformation into E. coli DH5a. Picked colony culture, rapid plasmid extraction for digestion, and expression in the temperature control conditions. The result was identified and the recombinant plasmid pBV / IL-2 / PEA was obtained. SDS-PAGE analysis of temperature-induced expression showed that there was a protein expression band at 51 000. Western blotting was performed with mouse anti-human IL-2 monoclonal antibody, and the expressed protein was designed as a chimeric protein. Conclusion This study established a method for preparing IL-2 / PEA chimeric protein, which can also be used to express other proteins.