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[重组人粒细胞-巨噬细胞集落刺激因子/白细胞介素-3融合蛋白的表达研究]

[Studies on the expression of the recombinant human GM-CSF/IL-3 fusion protein].

作者信息

Zhang Y, Qu X M, Yang S L

机构信息

Shanghai Research Center of Biotechnology, Chinese Academy of Sciences.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2000 May;16(3):316-9.

Abstract

A human granulocyte-macrophage colony stimulating factor (GM-CSF)/interleukin-3(IL-3) fusion gene with a short linker between the GM-CSF and IL-3 gene has been successfully constructed and expressed in E. coli under the control of T7 promoter. The recombinant fusion protein was expressed as inclusion bodies after the IPTG induction. The yield of the GM-CSF/IL-3 fusion protein was over 30% of the total cellular proteins. Western-blotting results showed that the fusion protein could specifically combined with GM-CSF antibody and IL-3 antibody. The biological activity was detected by the GM-CSF and IL-3 dependent cell line TF-1. After solubilizing with 8 mol/L urea and renaturing with dialysis against Tris. HCl solution, the refolded fusion protein showed obvious activities to maintain the growth of TF-1 cell.

摘要

一种在GM-CSF和IL-3基因之间带有短连接子的人粒细胞-巨噬细胞集落刺激因子(GM-CSF)/白细胞介素-3(IL-3)融合基因已成功构建,并在T7启动子控制下在大肠杆菌中表达。IPTG诱导后,重组融合蛋白以包涵体形式表达。GM-CSF/IL-3融合蛋白的产量超过细胞总蛋白的30%。Western印迹结果表明,融合蛋白能与GM-CSF抗体和IL-3抗体特异性结合。通过GM-CSF和IL-3依赖细胞系TF-1检测其生物学活性。用8mol/L尿素溶解并用Tris.HCl溶液透析复性后,复性的融合蛋白表现出明显的活性以维持TF-1细胞的生长。

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