兔早期胚胎发育相关新基因IFRG的克隆和表达

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。我们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IFRG cDNA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。

Cloning and expression of rabbit early embryo developmental new gene IFRG

Recent research has proved that interferons 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 (full 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. coli 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 41kDa and had immunity activity.