|
|||||
|
|
| 美洲大蠊变应原Cr PI的表达、纯化与免疫学特性鉴定 | |
| 引用本文： | 高波,刘志刚,邢苗,徐宏,罗时文,赖仞.美洲大蠊变应原Cr PI的表达、纯化与免疫学特性鉴定[J].昆虫学报,2005,48(1):13-17. |
| 作者姓名： | 高波 刘志刚 邢苗 徐宏 罗时文 赖仞 |
| 作者单位： | 1. 深圳大学生命科学学院,深圳,518060 2. 深圳大学生命科学学院,深圳,518060;深圳市微生物基因工程重点实验室,深圳,518060 3. 深圳市微生物基因工程重点实验室,深圳,518060 |
| 基金项目： | 国家自然科学基金项目 (3 9660 0 73 )，广东省科技重点项目 (2 0 0 3A3 0 80 5 0 2 )，深圳市科技计划资助项目 |
| 摘 要： | 以阳性噬菌体克隆为模板，通过PCR扩增出目的基因片段并克隆入T载体，经测序证实为美洲大
蠊Periplaneta americana变应原Cr PI后，将该基因亚克隆入表达载体pGEX-5X-1。美洲
大蠊变应原Cr PI在大肠杆菌中得到高效表达，但主要以包涵体形式存在于沉淀中。目的蛋白溶
于6 mol/L盐酸胍并经稀释复性后，经Glutathione SepharoseTM4B亲和层析，纯度达
90%以上。以蟑螂过敏病人血清进行免疫印迹检测，结果显示重组变应原具有良好的IgE结合活
性。 |
| 关 键 词： | 美洲大蠊 重组变应原Cr PI 蛋白表达 亲和层析 免疫印迹 |
| 文章编号： | 0454-6296(2005)01-0013-05 |
| 修稿时间： | 2003年12月29 |
Expression, purification and characterization of the American cockroach Cr PI allergen |
|
| GAO Bo,LIU Zhi-Gang,XING Miao,XU Hong,LUO Shi-Wen,LAI Ren.Expression, purification and characterization of the American cockroach Cr PI allergen[J].Acta Entomologica Sinica,2005,48(1):13-17. | |
| Authors: | GAO Bo LIU Zhi-Gang XING Miao XU Hong LUO Shi-Wen LAI Ren |
| Affiliation: | GAO Bo~1,LIU Zhi-Gang~ |
| Abstract: | Using the Cr PI clone from the λEXcell library as a template, the cDNA fragments were first generated by PCR techniques and then ligated into T vector. After being confirmed by DNA sequencing, the cDNA encoding the American cockroach Cr PI allergen was subcloned into pGEX-5X-1 and expressed as GST fusion protein in the form of inclusion bodies. After being dissolved in 6 mol/L guanidine hydrochloride and renatured with a simple dilution method, the proteins of target were purified to above 90% purity by affinity chromatography with Glutathione Sepharose 4B. Tested with sera from subjects allergic to cockroach, the recombinant allergen was shown to possess good IgE binding activity as determined by Western blotting. |
| Keywords: | American cockroach Periplaneta americana recombinant allergen Cr PI protein expression affinity chromatography Western blotting |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |
| 点击此处可从《昆虫学报》浏览原始摘要信息 | |
| 点击此处可从《昆虫学报》下载全文 | |
