【摘要】 IL-2是一种重要的免疫相关细胞因子,借助PCR去掉鸡IL-2基因片段(387bp)信号肽,连接到pPICZαA表达载体中,通过电转化转入Pichia酵母GS115菌株的感受态中,对筛选出来的阳性菌株以终浓度为1%的甲醇诱导表达,经SDS-PAGE及WesternBlot分析,结果表明酵母成功分泌表达了重组鸡IL-2,分子量为17.8kD。通过Ni-NTAHisBind柱层析,250mmol/L咪唑的洗脱液洗脱,得到纯度较高的重组鸡IL-2,淋巴细胞增殖试验证明,重组鸡IL-2具有明显的促进淋巴细胞增殖的生物活性,为进一步研究其免疫佐剂的作用提供了试验基础。更多还原

【Abstract】 Chicken IL-2 is a key cytokine for immunity. Removed the signal peptide by PCR, chIL-2 gene fragments (387 bp) was connected to the pPICZαA expression vector and transformed toPichia pastoris GS115 by electroporation. SDS-PAGE and Western Blot analysis showed that chIL-2 (17.8 kD) was successfully expressed and secreted by positive strains induced by 1% methanol. The chIL-2, which was purified by Ni-NTA His Bind column chromatography with 250 mmol/L imidazole elution buffer, had significant biological activity to promote lymphocyte proliferation, which provides the experimental basis for the further research on chIL-2 as a kind of immune adjuvant.更多还原