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目的 重组vMIPII在大肠杆菌中表达及其纯化。观察纯化的vMIPII蛋白对小鼠异体皮肤移植免疫排斥反应的抑制作用。方法 以 pET3 2a为vMIPII克隆基因的表达载体 ,以大肠杆菌BL2 1(DE3 ) plysS为表达菌 ,诱导表达出硫氧还蛋白 vMIPII的融合蛋白 (2 6× 10 3 D)。经金属离子亲和吸附、肠激酶消化以游离vMIPII、阳离子交换层析等步骤纯化目的蛋白。纯化的vMIPII从尾静脉连续 14d注入异体皮肤移植小鼠 (昆明鼠 /Balb/c)。结果 从 1L发酵菌液中可获得高纯度目的蛋白约 4mg。注射vMIPII的异体皮肤移植小鼠组移植皮存活时间较未注射对照组延长 7d。结论 纯化的重组vMIPII对小鼠异体皮肤移植免疫排斥反应有明显的抑制作用 ,能显著延长移植皮的存活时间
Objective Expression and purification of recombinant vMIPII in Escherichia. To observe the inhibitory effect of purified vMIPII protein on allogeneic skin graft rejection in mice. Methods pET3 2a was used as the expression vector for vMIPII cloning gene. The fusion protein of thioredoxin vMIPII (26 × 10 3 D) was induced by using E. coli BL21 (DE3) plysS as the expression vector. Adsorption of metal ions, enterokinase digestion to free vMIPII, cation exchange chromatography and other steps to purify the target protein. Purified vMIPII was injected into the allogeneic skin transplant mice (Kunming mouse / Balb / c) for 14 consecutive days from the tail vein. The results obtained from 1L fermentation broth high purity protein of about 4mg. Transplantation of allogeneic dermal transplantation mice injected with vMIPII prolonged the survival time of the grafted skin compared with the non-injected control group for 7 days. Conclusion The purified recombinant vMIPII can significantly inhibit the rejection of allogeneic skin grafts in mice and prolong the survival time of the grafted skin