Li Jiong, Cui Kaijun, Wen Jing, Zhao Zhiwei, Chen Ping, Tian Ling, Kan Bing, Wen Yanjun, Deng Hongxin, Fan Linyu, Wei Yuquan
State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2007 Aug;24(4):866-9.
A DNA fragment encoding human interleukin 4 was obtained by PCR from pORF-hIL4 plasmid. The amplified fragment was inserted into prokaryotic expression vector PQE60 and recombinant protein was expressed in E. Coli M15 by adding isopropyl-beta-D-thiogalactoside (IPTG). The hIL-4 protein was present as insoluble inclusion bodies in the bacterial extract. After denaturation of inclusion bodies with 5 mol/L guanidine hydrochloride, the supernate was diluted to get renaturized. Then dialysis and Ni chelating chromatography were used for purification. TF-1 proliferation assay of recombinant human interleukin 4 was performed, and then rhIL-4 was fit to be used for proliferation of human dendritic cells from monocyte in vitro.
通过聚合酶链反应（PCR）从pORF-hIL4质粒中获得编码人白细胞介素4的DNA片段。将扩增片段插入原核表达载体PQE60中，并通过添加异丙基-β-D-硫代半乳糖苷（IPTG）在大肠杆菌M15中表达重组蛋白。hIL-4蛋白以不溶性包涵体形式存在于细菌提取物中。用5mol/L盐酸胍使包涵体变性后，将上清液稀释以进行复性。然后通过透析和镍螯合层析进行纯化。对重组人白细胞介素4进行TF-1增殖测定，结果表明rhIL-4适合用于体外从单核细胞增殖人树突状细胞。
