重组人半乳凝集素-1的原核表达、纯化及生物活性检测  

Prokaryotic Expression,Purification and Biological Activity Determination of Human Galectin-1

作　　者：李晶[1] 贺丽清[1] 舒震[1] 侯建伟[1] 陈霖[1] 徐玉金[1] 张伟[1] 张英起[1] 

机构地区：[1]第四军医大学药学系生物制药学教研室,陕西西安710032

出　　处：《生物技术通讯》2010年第3期315-318,共4页Letters in Biotechnology

基　　金：国家自然科学基金(30972672;30801002)

摘　　要：目的:在大肠杆菌中表达半乳凝集素-1(galectin-1),并进行纯化及生物活性检测。方法:将人半乳凝集素-1基因克隆至带有His融合标签的原核表达载体pQE-30上,转化大肠杆菌M15,经IPTG诱导表达,表达产物经亲和层析纯化后,进行Western印迹鉴定,并用红细胞凝集试验检测其生物学活性。结果:双酶切鉴定和核苷酸序列测定表明重组表达质粒pQE-30-Galectin-1构建正确;重组蛋白的表达量约占菌体总蛋白的50%,主要以可溶形式表达,纯化后蛋白纯度达95%以上,且具有良好的红细胞凝集活性。结论:在大肠杆菌中表达了重组人半乳凝集素-1,且具有良好的生物活性。Objective：To express human galectin-1 in Escherichia coli and determine the biological activity of recombinant galectin-1.Methods：Galectin-1 gene was cloned into prokaryotic expression vector pQE-30 with His fusion labeling,and the constructed recombinant plasmid was transformed to E.coli M15 for expression under in-duction of IPTG.The expressed product was purified by affinity chromatography and identified by Western blot,then the biological activity was determined by hemagglutination test.Results：Restriction analysis and nucleotide sequencing proved that recombinant expression vector pQE-30-Galectin-1 was constructed correctly.The expressed product contained 50% of total somatic protein and mainly existed in a soluble form.After purification,the recom-binant galectin-1 reached a purity of more than 95% and showed good hemagglutinating activity.Conclusion：Re-combinant human galectin-1 was successfully expressed in E.coli and showed high biological activity.

关 键 词：重组人半乳凝集素-1 原核表达 纯化 红细胞凝集 

分 类 号：Q78[生物学—分子生物学]