林麝IL-2细胞因子的克隆、表达和纯化

用林麝（Moschus berezovskii）外周血单核细胞（peripheral blood mononuclear cells，PBMC）提取总RNA，RT-PCR扩增IL-2（interleukin-2，IL-2），连接pMD18-T载体后转化大肠杆菌，筛选阳性克隆并测序。然后将IL-2成熟肽编码序列连接到pGEX4T-1原核表达载体进行表达、纯化。结果表明林麝IL-2开放阅读框全长468bp，编码155个氨基酸。序列分析表明，林麝IL-2及其受体结合域与水牛的高度保守，而在翻译后修饰的预测活性位点上和水牛的有所不同。IFFG诱导林麝IL-2蛋白表达，得到约41kD的目标带；优化表达后，林麝IL-2表达含量达到了总蛋白量的25％。经GST亲和柱分离纯化，得到了约41kD的单一目标带，这说明林麝IL-2基因的原核表达成功。

Clone, Expression and Purification of Interleukin-2 from Moschus berezovskii

Moschus berezovskii IL-2 was cloned from Con-A stimulated peripheral blood mononuclear cells(PBMC),and ligated with pMD18-T vector,then transformed to Escherichia coli JM109.The positive recombinant was sequenced.The sequence encoding the mature peptide was ligated with pGEX4T-1 vector to express and purify the IL-2 protein.It showed that the length of M.berezovskii IL-2 was 468 bp,coding 155 amino acids.Sequence analysis of IL-2 showed that it had high homology with that of bubalus bubalis,except the difference of post-translational modification.After purified with GST,there was an aimed protein about 41kD.The result suggested that the Moschus berezovskii IL-2 was successfully cloned and expressed.