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用pKKH质粒在大肠杆菌中表达HuIFN-β

Expressing Human Fibroblast Interferon-β with Plasmid pKKH in Escherichia Colis
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摘要 应用DNA重组技术,将HuIFN-β基因插入到质粒pKKH的tac启动子下游,转化大肠杆菌JM101和JM103,经IPTG诱导,表达HuIFN-β,收集并裂解细菌,用Wish-VSV系统细胞病变抑制法检测生物学活性为2.18×108-8.7×108IU/L菌液。经初步纯化SDS-PAGE电泳可见分子量为20KD较纯的表达带。 Using recombinant DNA technology,HuIFM-β gene had been inserted into plasmid pKKH tac promoter downstream and transformated into Escherichia coli JM 101, JM 103 engineering bacteria and then cultured in LB culture medium, induced with IPTG and expressed rHuIFN-β.Collecting and lysising engineering bacteria,the biological activities of rHuIFN-β determinated by wish-VSV system CPE-restriction.The result was 2.18×10 8-8.7×10 8 IU/L.After primary purification, a 20 KD protein exprssing band can be seen in SDS-PAGE eletrophoretic figure.
出处 《微生物学免疫学进展》 1996年第2期42-45,共4页 Progress In Microbiology and Immunology
关键词 重组 人成纤维细胞 干扰素 rHuIFN-β 质粒 Recombination Human Fibroblast Interferon-β Tac promoter Gene expression
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