鹅IL-2成熟基因的表达及重组蛋白生物活性检测

Expression of GoIL-2 mature gene in E.coli

从PHA活化的东北白鹅外周血淋巴细胞中分离提取总RNA,利用反转录PCR技术扩增获得鹅IL-2(GoIL-2)基因,连接至克隆载体后进行测序鉴定,结果与预期大小一致。将去除信号肽的成熟基因亚克隆至原核表达载体,构建原核重组表达质粒并转化至大肠杆菌中进行诱导表达,用表达纯化的融合蛋白制备多克隆抗血清。此外,体外淋巴细胞增殖实验结果显示,鹅GoIL-2重组蛋白具有促进淋巴细胞增殖的活性,这为进一步研究GoIL-2的功能奠定了基础。

The goose interleukin-2 （GolL-2） cDNA was amplified by reverse transcription-polymerase chain reaction （RT- PCR） from peripheral blood lymphocyte of Northeast Goose that pre-treated with PHA. The PCR product was inserted into the cloning vector and confirmed by sequencing. The mature gene with signal peptide removed was subcloned to expression vector of procaryon, transformed into Escberichia eoli for expression. The expressed protein was purified and used to prepare polyclonal antiserum. In addition, the lymphopoiesis experiment in vitro showed that the rGolL-2 had the ability to promote lymphopoiesis.