北京鸭CD8α分子克隆与表达及其多克隆抗体制备  

Cloning,expression and polyclonal antibodies production of Peking duck CD8α

作　　者：廖定为[1] 唐秀山[1] 蒋一男[1] 张念之[1] 夏春[1] 

机构地区：[1]中国农业大学动物医学院,北京海淀100193

出　　处：《中国兽医杂志》2008年第6期5-7,共3页Chinese Journal of Veterinary Medicine

基　　金：病毒学国家重点实验室开放研究基金资助(2007No.8)

摘　　要：为了探讨鸭CTL免疫与病毒关系,本文用PCR技术从鸭cDNA文库中克隆了其CD8α基因(DuCD8α)。将DuCD8α成熟肽cDNA插入表达载体pQE30、构建了E.coliJM109(pQE30/DuCD8α)原核表达系。经诱导表达、蛋白纯化,制备了鼠抗DuCD8α多克隆抗体。结果表明DuCD8α成熟肽长645 bp,编码214个氨基酸。与已报道的DuCD8α在氨基酸水平上的同源性高达99.0%。SDS-PAGE证实重组DuCD8α(rDuCD8α)在JM109工程菌中的表达量可达15%。纯化的rDuCD8α分子量为26 kDa。ELISA结果显示鼠抗rDuCD8α多克隆抗体效价为1∶1 280 000。The duck CD8αchain （DuCD8α） was amplified by PCR method from the eDNA of Peking duck. The expression plasmid （pQE30/DuCD8α） containing the DuCD8α was constructed, and transformed into E. coli strain JM109, which was induced by IPTG to express recombinant protein. The expressed protein （rDuCD8α） was purified and analyzed by SDS-PAGE. In order to produce polyclonal antibodies, the Balb/c mice was immunized with the rDuCD8α. As results, the cloned DuCD8α mature peptide has 645 bp in length, encoding 214 amino acid residues. The homology with the reported DuCD8α is 99.0% at amino acid level. The SDS-PAGE analysis showed that the molecular weight is 26 kDa, and the mounts of the expressed rDuCD8αis 15.0% in total mass of bacterial protein by thin-layer scanner analysis. ELISA results revealed that the titer of anti-DuCD8α antibodies was about 1 ： 1 280 000.

关 键 词：北京鸭 CD8Α 表达 抗体 

分 类 号：Q789[生物学—分子生物学]