[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.
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