IL-1β的基因克隆及在原核中的表达

Cloning of IL-1β Gene and Expression in E.coli

从人外周血中分离出白细胞 ,提取其总RNA ,根据文献报道的IL 1 β的核苷酸序列合成 5′和 3′端引物 ,用RT PCR的方法获得了IL 1 β的基因cDNA ,并在大肠杆菌中获得了高效表达 ,表达量占全菌的 40 % ,并对表达产物进行了分离纯化和活性分析 ,获得了纯度大于 98%的样品 。

IL-1β cDNA was obtained by RT-PCR with the template of the total RNA extracted from leukocytes which was separated from human peripheral blood. 5′ and 3′ primers were synthesized according to literaturees reported sequence of IL-1β. IL-1β gene was highly expressed in E.coli and the expression level reached to about 40% of total bacteria proteins. Separation, purification and bioactivity analysis of the expressed products was performed. The purity of the final products reach more than 98%, and the culture solution of EL-4 cells induced by hIL-1β can promote the proliferation of CTLL-2 cells obviously.