鸡WDR72基因的克隆表达及其相应抗体的制备  

Cloning and Expression of Chicken WDR72 Gene andits Corresponding Antibody Preparation?

作　　者：李炳熠[1,2] 刘长国[1,2] 

机构地区：[1]浙江农林大学亚热带森林培育国家重点实验室培育基地,浙江临安311300 [2]浙江农林大学动物科技学院(筹),浙江临安311300

出　　处：《中国家禽》2012年第14期31-34,共4页China Poultry

基　　金：国家自然科学基金项目(30700567);浙江省自然科学基金项目(LY12C17002)

摘　　要：WDR72是课题组前期初步鉴定的蛋壳强度相关基因,它是个新基因。本文首先参考GenBank中原鸡(G.gallus)的WDR72基因序列设计3对引物,从仙居鸡子宫组织的cDNA中克隆WDR72基因,并构建家鸡WDR72编码基因的pEGFP-N1真核表达重组质粒;其次,将所克隆的WDR72基因的3′端编码序列构建到原核表达质粒pET-28b,然后利用0.4mmol/L的IPTG于37℃诱导表达WDR72的C端肽链;最后利用500mmol/L的咪唑纯化该肽链,并免疫大白兔制备wdr72特异性多抗。上述工作将为深入研究WDR72基因的生物学特性及其生化功能提供基础。Our group has previously identified a novel gene, WDR72,which is associated with eggshell strength. In present paper, firstly, 3 pairs of primers were designed according to the sequence of gallus （G.gallus） in GenBank to chine WDR72 gene from the cDNA of Xianju chicken, and the eukaryotic expression recombinant plasmid pEGFP-N1 of WDR72 was made; secondly, the 3＇ sequence of WDR72 gene was constructed into prokaryotic expression plasmid pET-28b,then 0.4mmol/L IPTG was used to induce the recombinant protein of WDR72-C;finally,through purification with 500 mmol/L imidazole, the polypeptide chain was obtained and then was further used to immunize white rabbit for WDR72 polyclonal antibody. This study will facilitate the in-depth expression characteristics and biochemical function of WDR72 study.

关 键 词：鸡 WDR72 基因克隆 原核表达与纯化 

分 类 号：Q943.2[生物学—植物学]