摘 要：半乳糖凝集素-3 (galectin-3, Gal-3)与肿瘤的发生发展密切相关，已有文献表明其羧基端galectin-3C (Gal-3C)具有一定的肿瘤抑制作用。为了获得Gal-3C的抗血清，实验以已有的重组质粒pGM-Gal-3为模板，利用PCR方法扩增中华大蟾蜍Gal-3C基因片段，并将其插入到表达载体pET28b上，构建中华大蟾蜍Gal-3C的原核表达载体pET28b-Gal-3C；将构建好的重组质粒pET28b-Gal-3C转入感受态细胞E. coli BL21 (DE3)中，用IPTG诱导蛋白质表达，并通过SDS-PAGE和Western-blot检测目的蛋白Gal-3C的表达；以纯化的重组蛋白Gal-3C免疫小白鼠，制备鼠抗中华大蟾蜍Gal-3C的抗血清，用Western-blot检测抗血清的特异性及其效价。结果显示成功构建了中华大蟾蜍pET28b-Gal-3C重组质粒，并诱导得到了目的蛋白的表达，而且将表达产物免疫小鼠后获得了特异性良好的抗中华大蟾蜍Gal-3C的抗血清。以上结果为抗肿瘤药物的研究和发展奠定了基础。

Abstract: Galectin-3 (Gal-3) is closely related to the genesis and development of tumor. Its derivative galectin-3C (Gal-3C) has great inhibitory effect on tumor. In order to prepare the antiserum of Gal-3C, Gal-3C fragment was amplified by PCR, using the existing recombinant plasmid pGM-Gal-3 as template, and inserted into expression vector pET28b to construct pET28b-Gal-3C, the prokaryotic expression vector of Chinese toad (Bufo gargarizans) Gal-3C. The recombinant plasmid was transferred into E. coli competent cells BL21 (DE3). The expression of recombinant Gal-3C was induced by IPTG and determined by SDS-PAGE and Western-blot assays. Then mice were immunized with the purified recombinant Gal-3C to prepare murine antiserum against Gal-3C. The specificity of the antiserum was determined by Western-blot. The results showed that pET28b-Gal-3C was successfully constructed and that recombinant Gal-3C and its specific antiserum were obtained. The current study builds a foundation for research and development of anti-cancer drugs.