TGF-β信号家族在生物体生命活动中具有多种发育和生理功能。研究该信号家族在生物体内的表达模式具有重要的意义。gbb是TGF-β信号成员bmp7的果蝇同源基因,为了进一步研究gbb对胚胎发育的影响, 本文采用DNA免疫技术制备得到果蝇GBB多克隆抗体。首先提取果蝇总RNA, 反转录得到cDNA文库,以该文库为模板克隆出gbb基因编码区序列, 构建成真核表达质粒pCAGGS-P7/gbb。采用DNA免疫技术免疫BALB/c小鼠, 取心血分离血清制备了GBB多克隆抗体。免疫印迹试验表明制备的GBB多克隆抗体有效,果蝇胚胎免疫荧光分析表明GBB在果蝇胚胎发育0~9 h无明显表达, 9 h之后开始高效表达。

The TGF-β signaling plays multiple developmental and physiological functions in life. It is important to study the expression pattern of this signal family in vivo. gbb is a homolog gene of bmp7 in Drosophila. In order to study the effect of TGF-β signaling on embryonic development, a polyclonal antibody against Drosophila GBB was prepared. The cDNA sequence of the gbb was cloned (from the library) by cDNA library of Drosophila, and the eukaryotic expression plasmid pCAGGS-P7/gbb was constructed. The BALB/c mice were immunized with DNA immunoassay, and the polyclonal antibody against GBB was prepared by taking serum. The antibody was found to be effective by WB. Through immunofluorescence of Drosophila embryos, it was shown that GBB has no obvious expression at 0~9 h, however, highly expressed signal may be seen after 9 h.