目的:表达具备生物活性的重组小鼠Wnt-3a信号蛋白.方法:应用脂质体转染试剂将重组真核表达载体pSecTag2/HygroB-Wnt3a转染并筛选稳定表达的NIH3T3细胞,Western Blot鉴定重组Wnt-3a蛋白的表达,并对Wnt3a/NIH3T3细胞的融合密度及抗凋亡能力给予检测.结果:Wnt-3a信号蛋白在Wnt3a/NIH3T3细胞中获得稳定表达,Wnt-3a信号蛋白能够明显提高NIH3T3细胞的融合密度及抗凋亡能力.结论在NIH3T3细胞中表达的重组Wnt-3a信号蛋白具备生物活性.
AIM: To express recombinant mouse Wnt-3a signal protein with biological activity. METHODS: The recombinant eukaryotic expression plasmid, pSecTag2/Hygro B-Wnt3a, was transfected into NIH3T3 cells by liposome, then the expressed protein was detected by Western Blot. The saturation density and the capability of suppressing apoptosis in Wnt3a/NIH3T3 cells were evaluated. RESULTS: The Wnt-3a signal protein was stably expressed in Wnt3a/NIH3T3 cells. The saturation density and the capability of suppressing apoptosis in Wnt3a/NIH3T3 cells were markedly increased. CONCLOUSION: We successfully construct the recombinant Wnt-3a signal protein with biological activity.