|
|||||
|
|
| 橡胶延长因子基因的原核表达及其多克隆抗体的制备 | |
| 引用本文： | 陈章权,吴坤鑫,梁晓东,黄震,陆田田.橡胶延长因子基因的原核表达及其多克隆抗体的制备[J].安徽农业科学,2008,36(29). |
| 作者姓名： | 陈章权 吴坤鑫 梁晓东 黄震 陆田田 |
| 作者单位： | 1. 广东医学院临床免疫学教研室,广东湛江,524023;中国热带农业科学院热带生物技术研究所,海南海口,571101 2. 中国热带农业科学院热带生物技术研究所,海南海口,571101 3. 广东医学院临床免疫学教研室,广东湛江,524023 |
| 基金项目： | 国家自然科学基金 ， 中央级公益性科研院所基本科研业务费专项资金资助项目 |
| 摘 要： | 目的]制备橡胶延长因子重组蛋白及其多克隆抗体。方法]用RT-PCR法扩增橡胶延长因子(rubber elongation factor,REF)编码区基因,将其克隆到原核表达载体pDEST17中,转化大肠杆菌BL21-AI,用L-阿拉伯糖诱导重组蛋白的表达,并采用亲和层析法纯化重组蛋白,以之为免疫原免疫小鼠制备多克隆抗体,并用ELISA和W estern blot对抗体效价和特异性进行鉴定。结果]高效表达和纯化了橡胶延长因子重组蛋白,用其免疫家小鼠,获得了高效价的特异性多克隆抗体,W estern blot检测显示该抗体可识别胶乳中的天然橡胶延长因子。结论]成功获得橡胶延长因子重组蛋白,制备了效价高、特异性好的鼠抗橡胶延长因子抗体,为进一步橡胶延长因子及其他橡胶粒子膜蛋白的生物学功能研究奠定了基础。 |
| 关 键 词： | 橡胶延长因子 基因 表达 抗体 制备 |
Prokaryotic Expression of Rubber Elongation Factor Gene and Preparation of Its Polyclonal Antibody |
|
| Abstract: | Objective] The aim of this study was to prepare the recombination protein of rubber elongation factor and its polyclonal antibodies.Method] The encoding gene of rubber elongation factor(REF) was amplified by RT-PCR,and cloned into the prokaryotic expression vector pDEST17 to transform into Escherichia coil BI2I-AI.The recombinant protein induced by L-Arabinose was purified by the affinity chromatography.As the immunogen,the recombination protein was used to immunize mice for preparing polyclonal antibodies,while ELISA and Western blot hybridization were used to detect the titers and specificity.Result] The purified recombination protein of rubber elongation factor with high expression was used to immunize house mice for preparing polyclonal antibodies with high titer and specificity.The Western blot hybridization showed that the antibody could recognize the natural rubber elongation factor from latex.Conclusion] The recombination protein of rubber elongation factor is successfully obtained and the mouse anti rubber elongation factor antibody with high titer and specificity is prepared,which lays a basis for further studies on biological functions of rubber elongation factor and other membrane proteins in rubber particles. |
| Keywords: | Rubber elongation factor Gene Expression Antibody Preparation |
| 本文献已被 维普 万方数据 等数据库收录！ | |