猪IL-2的原核表达及其促淋巴细胞增殖活性研究

Prokaryotic expression of porcine IL-2 and its stimulating activity for lymphocyte proliferation

为研究猪IL-2作为免疫佐剂的可行性,用IPTG诱导含有猪IL-2基因的BL21(DE3)LysS工程菌。目的蛋白经超声波破碎、洗涤、变性溶解后,用固化金属亲和层析法纯化后复性。MTT法对复性的重组蛋白的生物学活性进行分析。结果显示,相同浓度复性的IL-2与未复性的IL-2比较,刺激淋巴细胞体外增殖的能力差异极显著(P<0.01)。说明复性的重组猪IL-2具有良好的刺激淋巴细胞体外增殖的活性,在作为免疫佐剂方面具有潜在的应用价值。

To explore the feasibility of porcine interleulein-2 （plL-2） as immunoadjuvant, Escherichia coli BL21 （DE3） LysS with the gene encoding plL-2 was induced by IPTG. The expressed protein was in inclusion body form. The dissolved inclusion body was purified by Immoble Metal Affinity Chromatography （IMAC） and renaturated. The lymphocyte proliferation activity of the renatured protein IL-2 was examined using MTT colorimetric analysis. The MTT result showed that the renatured protein IL-2 had good stimulating activity for lymphocyte proliferation, indicating that pIL-2 may have potential as immunoadjuvant.