带有组氨酸标签的蛇毒基因在毕赤酵母中的表达

Expression of A Snake Venom Gene with Histidine Tag in Pichia pastoris

[目的]通过巴斯德毕赤酵母(Pichia pastoris)表达带有组氨酸标签的蛇毒基因。[方法]利用基因重组技术,以海蛇基因为材料,通过PCR技术在N末端添加一个六聚组氨酸纯化标签,并将重组质粒导入菌株GS115,用1%甲醇诱导后,分泌表达了重组蛋白。[结果]测序结果显示该标签已成功插入,SDS-PAGE检测到分子量为28.5 kD的目的蛋白。[结论]成功表达了带有组氨酸标签的蛇毒蛋白,其具有良好的降纤活性。

[Objective] A snake venom gene with Histidine Tag was expressed through Pichia pastoris.[Method] Using the gene recombination technology,with the sea snake as the material,a 6×His Tag was inserted into the N end of the snake venom gene through the PCR technology and the restructured plasmid was transferred into the GS115 strains,after which was induced by 1% methanol,the recombinant protein was secreted and expressed.[Result] The sequencing results revealed that DNA 6×His Tag was accurately inserted into the expression vector,the purpose protein with the molecular weight of 28.5 was detected by SDS-PAGE.[Conclusion] The snake venom gene with Histidine Tag was expressed successfully and it had good defibre activity.