成纤维细胞生长因子与单核-巨噬细胞集落刺激因子融合蛋白在大肠杆菌中的表达

Expression of Recombinant Fusion Protein Basic Fibroblast Growth Factor and Granulocyte-Macrophage Colony Stimulating Factor in E coli

摘要:

为获得防治动脉粥样硬化和再狭窄的新型基因产品,应用基因工程技术将碱性成纤维细胞生长因子和单核─巨噬细胞集落刺激因子基因克隆到pGEM-3Zf(+)载体的EcoRI和BamHI位点上,重组成融合基因,亚克隆到pBV220表达载体上,并在大肠杆菌DHα中进行表达,然后用十二烷基磺酸钠─聚丙烯酸胺凝胶电泳和Westernblot进行分析。结果发现,融合蛋白可获得表达,且具有成纤维细胞生长因子与单核─巨噬细胞集落刺激因子的免疫学活性。这为进一步研究该融合蛋白的功能及防治动脉粥样硬化和动脉再狭窄提供一种新的基因产品。

Abstract:

Aim: This research has rnade a new gene product for the treatment of atherosclerosis and arterial stenosis.Methods: By rneans of gene engineering technology,basic fibroblast growth factor (bFGF) cDNA and granulocyte macrophage colony stimulating factor (GMCSF) cDNA were cloned into the vector pGEM-3Zf (+ ) at the EcoR I and BamHI sites. Subsequntly subcloned into the expression vector pBV220 of the EcoR I and BamH I sites. The bFGF-GM-CSF was expressed in E coli confirmed by SDS-PAGE and Western-bolt.Results: The recombinant fusion protein was expressed successfully in E coli. They have immunological activities of bath bFGF and GM-CSF.Conclusion: The recombinant fusion protein may have biological activities of GM-CSF.