摘要
zfol是斑马鱼的一种母源凝集素基因。从斑马鱼胚胎中提取RNA,应用RT-PCR从斑马鱼中得到zfol的基因片段,重组入原核表达载体pEASY-E2中得到重组质粒,并测序鉴定其序列的正确性,将重组质粒转化到大肠杆菌BL21中诱导表达,并纯化蛋白,利用纯化的重组蛋白为抗原免疫兔子,制备斑马鱼母源凝集素Zfol蛋白的多克隆抗体,利用Western-blot方法检测抗体特异性,得到特异的多克隆抗体,为进一步研究母源凝集素基因zfol的功能奠定了基础。
Zfol is a maternal lectin gene of zebrafish.In this study,the total RNA was isolated from the zebrafish.The zfol gene fragment was amplified by RT-PCR and subcloned into a prokaryotic expression vector pEASY-E2.Then transformed the recombinant plasmid into E.coli BL21.The recombinant protein was purified to prepare polyclonal antibody.The purified recombinant protein was used to immunize rabbits for production of polyclonal antibodies against the protein of zfol gene and generated the polyclonal antibody with identification by Western blot method.The specific polyclonal antibody was obtained,and it would be used to the functional assay.
出处
《安徽农学通报》
2013年第6期14-16,共3页
Anhui Agricultural Science Bulletin
基金
母源凝集素ZfOL在斑马鱼胚胎发育早期的免疫保护作用研究
关键词
斑马鱼
zfol
原核表达
多克隆抗体
Zebrafish
Zfol
Prokaryotic expression
Polyclonal antibody
