High-level expression of a bioengineered, cysteine-free hepatocyte-stimulating factor (interleukin 6)-like protein.
High-level expression of a bioengineered, cysteine-free hepatocyte-stimulating factor (interleukin 6)-like protein.
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高水平表达生物工程、不含半胱氨酸的肝细胞刺激因子（白细胞介素 6）样蛋白。
DOI：
10.1073/pnas.85.24.9426
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发表时间：
1988
影响因子：
11.1
通讯作者：
Fowlkes,DM
中科院分区：
文献类型：
--
作者：
Jambou,RC;Snouwaert,JN;Bishop,GA;Stebbins,JR;Frelinger,JA;Fowlkes,DM
肝细胞刺激因子、干扰素- β 2、b细胞刺激因子2和杂交瘤/浆细胞瘤生长因子是相同的蛋白质，目前被称为白细胞介素6 （IL-6）。利用合成寡核苷酸技术，基于人IL-6 cDNA序列构建了具有生物活性的重组IL-6 （IL-6）基因。该合成基因编码一种无半胱氨酸的生物工程rIL-6蛋白，该蛋白在大肠杆菌中作为三方融合蛋白高水平表达。融合蛋白与胶原酶的裂解释放一个23 kda的rIL-6蛋白，可以很容易地纯化到均匀性。我们发现，IL-6蛋白显示出一系列类似于天然人IL-6的生物活性，如其能够(i)保护细胞免受病毒感染，（ii）刺激大鼠FAZA 967细胞纤维蛋白原的合成，以及（iii）诱导B细胞的终末分化，导致免疫球蛋白分泌增加。
Hepatocyte-stimulating factor, interferon-beta 2, B-cell stimulation factor 2, and hybridoma/plasmacytoma growth factor are identical proteins presently referred to as interleukin 6 (IL-6). Through the use of synthetic oligonucleotide technology, we have constructed a biologically active recombinant IL-6 (rIL-6) gene based on the sequence of a human IL-6 cDNA. The synthetic gene encodes a cysteine-free, bioengineered rIL-6 protein that is expressed at high levels in Escherichia coli as a tripartite fusion protein. Cleavage of the fusion protein with collagenase releases a 23-kDa rIL-6 protein that can be easily purified to homogeneity. We show that the rIL-6 protein displays a range of biological activities similar to those of natural human IL-6, as demonstrated by its ability to (i) protect cells from viral infection, (ii) stimulate the synthesis of fibrinogen in rat FAZA 967 cells, and (iii) induce the terminal differentiation of B cells, resulting in elevated secretion of immunoglobulin.