安徽农学通报
安徽農學通報
안휘농학통보
AUHUI AGRICULTURAL SCIENCE BULLETIN
2013年
6期
14-16
,共3页
斑马鱼%zfol%原核表达%多克隆抗体
斑馬魚%zfol%原覈錶達%多剋隆抗體
반마어%zfol%원핵표체%다극륭항체
Zebrafish%Zfol%Prokaryotic expression%Polyclonal antibody
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是斑馬魚的一種母源凝集素基因.從斑馬魚胚胎中提取RNA,應用RT-PCR從斑馬魚中得到zfol的基因片段,重組入原覈錶達載體pEASY-E2中得到重組質粒,併測序鑒定其序列的正確性,將重組質粒轉化到大腸桿菌BL21中誘導錶達,併純化蛋白,利用純化的重組蛋白為抗原免疫兔子,製備斑馬魚母源凝集素Zfol蛋白的多剋隆抗體,利用Western-blot方法檢測抗體特異性,得到特異的多剋隆抗體,為進一步研究母源凝集素基因zfol的功能奠定瞭基礎.
zfol시반마어적일충모원응집소기인.종반마어배태중제취RNA,응용RT-PCR종반마어중득도zfol적기인편단,중조입원핵표체재체pEASY-E2중득도중조질립,병측서감정기서렬적정학성,장중조질립전화도대장간균BL21중유도표체,병순화단백,이용순화적중조단백위항원면역토자,제비반마어모원응집소Zfol단백적다극륭항체,이용Western-blot방법검측항체특이성,득도특이적다극륭항체,위진일보연구모원응집소기인zfol적공능전정료기출.
@@@@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.
