利用DNA免疫技术制备果蝇GBB多克隆抗体及其表达模式的分析

王 璐,查浩浩,江志钢

激光生物学报 ›› 2019, Vol. 28 ›› Issue (5) : 458-462.

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激光生物学报 ›› 2019, Vol. 28 ›› Issue (5) : 458-462.
研究论文

利用DNA免疫技术制备果蝇GBB多克隆抗体及其表达模式的分析

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Preparation of Polyclonal Antibody Against GBB by DNA Immunization and Preliminary Study on Its Expression

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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。

Abstract

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.

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王 璐,查浩浩,江志钢. 利用DNA免疫技术制备果蝇GBB多克隆抗体及其表达模式的分析[J]. 激光生物学报. 2019, 28(5): 458-462
WANG Lu, ZHA Haohao, JIANG Zhigang. Preparation of Polyclonal Antibody Against GBB by DNA Immunization and Preliminary Study on Its Expression[J]. Acta Laser Biology Sinica. 2019, 28(5): 458-462

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