Abstract: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.
引用本文:
刘怡君, 贾宇坤, 袁进强, 蒋桂瑾, 岳继萍, 李玉兰, 杨仙玉. 中华大蟾蜍重组Gal-3C的原核表达、纯化及抗血清制备[J]. 生命科学研究, 2016, 20(6): 497-501.
LIU Yi-Jun, JIA Yu-Kun, YUAN Jin-Qiang, JIANG Gui-Jin, YUE Ji-Ping, LI Yu-Lan, YANG Xian-Yu. Prokaryotic Expression, Purification and Antiserum Preparation of Recombinant Gal-3C of Bufo gargarizans. Life Science Research, 2016, 20(6): 497-501.