| 重组人乙酰肝素酶在大肠杆菌中的表达、纯化 |
| |
| 引用本文： | 高红伟,田曙光,崔真源,李素波,檀英霞,王璇琳,冉菘余,宫锋.重组人乙酰肝素酶在大肠杆菌中的表达、纯化[J].军事医学科学院院刊,2009,33(5):434-437. |
| |
| 作者姓名： | 高红伟 田曙光 崔真源 李素波 檀英霞 王璇琳 冉菘余 宫锋 |
| |
| 作者单位： | 军事医学科学院野战输血研究所,北京,100850 |
| |
| 基金项目： | 科技部支撑计划重大项目"常见恶性肿瘤分子标志的临床评价" |
| |
| 摘 要： | 目的：表达纯化重组的人乙酰肝素酶（heparanase,HPA）。方法：以表达乙酰肝素酶全长质粒pcDNA3.1-HPA为模板,扩增不包括信号肽的乙酰肝素酶片段,将其插入pET-28a（＋）载体,转化到大肠杆菌BL21（plysS,DE3）中,在起始诱导菌密度、诱导温度、诱导时间、诱导剂浓度4个方面对诱导条件进行了优化,实现了重组人乙酰肝素酶的可溶性表达。采用HisTrapTMcrude亲和层析对目的蛋白进行了初步纯化。结果：得到了初步纯化的HPA蛋白,Western印迹证实表达产物可被乙酰肝素酶抗体和His标签抗体识别,证明表达产物具有乙酰肝素酶的免疫学特性。结论：表达纯化的人HPA蛋白为特异性单抗的制备和鉴定提供了实验材料。
|
| 关 键 词： | 乙酰肝素酶 大肠杆菌 表达优化 蛋白纯化 |
Expression and purification of recombinant human heparanase in Escherichia coli |
| |
| GAO Hong-Wei,TIAN Shu-Guang,CUI Zhen-Yuan,LI Su-Bo,TAN Ying-Xia,WANG Xuan-Lin,RAN Song-Yu,GONG Feng.Expression and purification of recombinant human heparanase in Escherichia coli[J].Bulletin of the Academy of Military Medical Sciences,2009,33(5):434-437. |
| |
| Authors: | GAO Hong-Wei TIAN Shu-Guang CUI Zhen-Yuan LI Su-Bo TAN Ying-Xia WANG Xuan-Lin RAN Song-Yu GONG Feng |
| |
| Institution: | (Beijing Institute of Transfusion Medicine, Beijing 100850, China) |
| |
| Abstract: | Objective：To express,purify and identify the recombinant human heparanase. Methods：The cDNA of truncated human heparanase （without signal peptide） was cloned by PCR using pcDNA3.1-HPA as template and subcloned into pET-28a （ ＋ ） vector. The expression plasmid was transformed into Escherichia coli BI2,1 （plysS, DE3 ） to produce soluble protein. The protein was purified by means of affinity chromatography. Results： The result showed that the expressed protein reacted with anti-His antibody and anti-beparanase antibody respectively. Conclusion：We obtain a soluble expression protein in prokaryotic expression system, which has immunological characteristics of heparanase. |
| |
| Keywords: | heparanase Escherichia coli expression optimization protein purification |
| 本文献已被 维普 万方数据 等数据库收录！ |
|
