摘要
为了进一步研究DJ-1蛋白在J亚群禽白血病病毒感染中的生物学作用,本研究从DF-1细胞的cDNA中扩增DJ-1基因编码区,并克隆入pET-32a载体进行原核表达。结果表明:表达的重组蛋白以可溶性形式存在,最佳表达条件为IPTG终浓度为0.4mmol/L,37℃诱导3h;ELISA和Western结果证明融合蛋白免疫BALB/c小鼠后获得的多抗血清具有良好的特异性和反应性。本试验中鸡源DJ-1蛋白及小鼠多抗血清的获得,为后期DJ-1蛋白功能性研究提供了实验材料。
To investigate the function of DJ-1 gene of chicken , a panel of polyclonal antibodies against DJ-1 was needed. In this paper , the CDS region of DJ-1 from cDNA of DF-1 cells were amplified by PCR and cloned it into pET-32a for prokaryotic expression. The BALB/c mice were immunized with the expressed recombinant protein. The results showed that the recombinant DJ-1 fusion protein was abundant and soluble under 3 h induced with 0.4mmol/L IPTG at 37 ℃. ELISA and Western blot results demonstrated that the anti-serum from mice specifically reacted with the recombinant DJ-1. The sera will support the further study of DJ-1 function in the avian leukosis virus subgroup J infection.
出处
《中国家禽》
北大核心
2012年第11期32-35,共4页
China Poultry
基金
联合基金重点项目(U0831002)
教育部创新团队(IRT0978)
江苏省优势学科项目
