人单核细胞趋化蛋白-1在大肠杆菌中的高效表达及其生物学活性

High Level Expression of Human Monocyte Chemoattractant Protein-1 in Escherichia Coli and its Biological Activity

利用PCR和DNA重组技术，将人单核细胞趋化蛋白-1（MCP-1）cDNA克隆在表达载体pGEX－2T中，表达产物GST／MCP－1占菌体总蛋白的50％，且以无活性的包涵体形式存在。经变复性，GlutathioneSepharose4B亲合柱层析纯化，融合蛋白达到电泳纯，且具有MCP－1趋化活性。体内外抗肿瘤试验结果表明，GST／MCP－1激活单核细胞和淋巴细胞后可抑制肿瘤细胞生长，提示MCP－1可能成为肿瘤辅助治疗的一个候选药物。

With PCR and DNA recombination technology, a cDNA encoding human monocytechemoattractant protein-1 (MCP- ) was cloned into the expression vector PGEX-2T. The GST/MCP-1 fusion protein was overexpressed in E.coli and accounted for about 50% of total cellularproteins. The GST/MCP-1 were in inclusion bodies. After denaturation and renatUfation in vitro,the renatured products were purified to be electrophoretically pure by affinity chromatography onglutathione Sepharose 4B. The purified product retains higher biological activities. The vitro and invivo antitumor effect showed that GST/MCP-1 could activate monocytes and lymphocytes to inhibitthe growth of a lung adenocarcinoma cell line A549. These results suggest that MCP-1 may becomea candidate for cancer immunotheraphy.