学术
摘 要:
为了获得鸡CD8α/β蛋白及其多克隆抗体,进一步研究其结构与功能,本实验从三黄鸡cDNA文库中克隆了其CD8α/β胞外区片段,构建了pQE30/ChCD8α/β原核表达系统,并经诱导表达、亲和层析纯化,获得了纯化的重组蛋白（rChCD8α/β）,然后用rChCD8α/β分别免疫Balb/c小鼠制备了其多克隆抗体。结果表明：本实验克隆的ChCD8α长483 bp、编码161个氨基酸、可在E.coliJM109中高效表达,蛋白质分子质量为24.0 ku,表达量占菌体蛋白量的25.0%,由该蛋白质所制备的多克隆抗体抗rChCD8α的ELISA效价为1∶1 024 000;另外,克隆的ChCD8β长447 bp,编码149个氨基酸,蛋白质的分子质量为18.5 ku,表达量占菌体蛋白量的20.0%,抗rChCD8β多克隆抗体ELISA效价为1∶64 000。本实验所得鸡CD8α/β的多克隆抗体将用于四聚体研究。[著者文摘]
文章出处:
《中国农业大学学报》-2007年12卷5期 -5-9页
栏目信息:
文献标识码:
A
文章编号:
1007-4333（2007）05-0005-01
Cloning, expression and production of polyclonal antibodies in extracellular regions of chicken CD8α /β
Tang Xiushan, Wang Lei, Liao Dingwei, Jiang Yi＇nan, Xia Chun （College of Veterinary Medicine, China Agricultural University, Beijing 100094, China）
Abstract:
The aim of this research was to prepare proteins and polyclonal antibodies for further study of the structure and function of chicken CD8α /β. The extracellular regions of CD8α /β chains were amplified by PCR and expression systems （pQE30/ChCD8α /β） were constructed and induced. The recombinant protein （rChCD8α /β） was purified to immunize the Balb/c mice. The resulting cloned ChCD8α had 483 bp in length, encoding 161 amino acid residues. SDS-PAGE analysis showed that the molecular weight was 24.0 ku, and the mounts of the recombinant protein was 2.5.0% of total mass of bacterial protein. ELISA analysis showed that the titer of polycIonal anti-ChCD8α was about 1 ： 1 024 000. The cloned ChCD8α had 447 bp in length, encoding 149 amino acid residues. The SDS-PAGE analysis showed that the molecular weight was 18.5 ku, and the mounts of the recombinant protein was 20.0% of total mass of bacterial protein. ELISA analysis showed that the titer of polycIonal anti-ChCD8α was about 1：64 000. Antibodies provided from the research can be applied to the further study of tetramer.[著者文摘]
Key words:
chicken;CD8α; CD8β; prokaryotic expression; polycional antibody
基金资助:
国家自然科学基金资助项目（30671566）
更多>>免费文章
cqvip.com