兔早期胚胎发育相关新基因IFRG的克隆和表达  

Cloning and Expression of Rabbit Early Embryo Developmental New Gene IFRG

作　　者：齐冰[1] 何新[1] 陈清轩[1] 

机构地区：[1]中国科学院遗传与发育生物学研究所

出　　处：《中国生物工程杂志》2006年第6期55-58,共4页China Biotechnology

基　　金：国家"973"计划资助项目(G200016107)

摘　　要：近来的研究表明干扰素在哺乳动物早期胚胎发育中有重要的作用。首次克隆了兔早期胚胎发育相关新基因IFRG(干扰素应答基因)的全长cDNA序列(AJ584672),根据该cDNA序列以兔卵巢cDNA为模板,经PCR扩增后克隆了兔IFRGcDNA的完整开放阅读框(396bp)。将其克隆到原核表达载体pGEX-4T-2上,在大肠杆菌BL21中进行了GST-IFRG融合蛋白的表达。经IPTG诱导培养后,SDS-PAGE电泳检测结果显示在41kDa处有特异性表达蛋白,回收融合蛋白作为抗原免疫小鼠,制备多克隆抗体,通过Western杂交表明该融合蛋白具有生物免疫活性。Recent reseach has proved that interfercons play very important role in early embryo development. The full length cDNA （AJ584672） of rabbit early embryo developmental new gene IFRG （ interferon responsive gene） was cloned. According to rabbit IFRG cDNA, intact open reading frame （ hall length was 396bp） was amplified using rabbit ovary cDNA as template. The purified rabbit IFRG cDNA was inserted into pGEX-4T-2 vector to construct the prokaryotic expression vector. IFRG protein was expressed in E. coil in the form of glutathione-S-transferase （GST） fusion protein. The recombinant expression plasmid was transformed into BL21 and gene expression was induced by IPTG. Expression protein were detected by SDS-PAGE and confirmed by Western blotting analysis, which showed the molecular weight of the protein was 41 kDa and had immunity activity.

关 键 词：家兔 IFRG RT-PCR 融合蛋白 WESTERN 杂交 

分 类 号：Q786[生物学—分子生物学]