| 摘要: |
| 克隆了p27蛋白基因并构建了重组表达质粒pET28a-p27,该质粒在大肠杆菌BL21中经IPTG诱导后获得了可溶性表达蛋白,经辣根过氧化物酶标记的兔抗p27抗体进行Western blotting检测结果为阳性。后经金属螯和层析纯化获得纯度约为95%的蛋白样品,用纯化后的p27蛋白直接作为抗原免疫家兔产生抗血清,经ELISA检测抗体效价达1：25600。结果证明,成功地制备了特异性的p27多克隆抗体。 |
| 关键词: 禽白血病病毒 P27蛋白 原核表达 蛋白印记 多克隆抗体 |
| DOI： |
| 投稿时间：2013-10-14修订日期：2013-10-30 |
| 基金项目: |
|
| Expression of p27 protein of Avian leukosis virus in Escherichia coli and preparation of its polyclonal antibody |
|
| (China Institute of Veterinary Drug Control,Beijing,100081) |
| Abstract: |
| p27 protein gene from Avian leukosis virus was cloned and constructed with plasmid pET28a.The recombinant protein p27 was expressed in an E.coli strain BL21(DE3) with IPTG induction and detected by Western blotting with horseradish peroxidase labeled rabbit-anti-p27 antibodies. After that,the protein was purified to above 95% of purity by affinity chromatography with Ni-NTA agarose column.The anti-sera against p27 protein was obtained by immunizing rabbit with the purified recombinant p27 protein .The specificity of polyclonal antibody was about 1：256000 after detected by ELISA. Thus the high specific polyclonal antibody of p27 protein was proved to be prepared． |
| Key words: Avian leukosis virus p27 protein expression Western blotting polyclonal antibody |
