| 带有组氨酸标签的蛇毒基因在毕赤酵母中的表达 |
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| 引用本文： | 黎卓键.带有组氨酸标签的蛇毒基因在毕赤酵母中的表达[J].安徽农业科学,2011,39(31):19054-19056. |
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| 作者姓名： | 黎卓键 |
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| 作者单位： | 暨南大学生物工程研究所,广东广州,510632 |
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| 基金项目： | 广东省教育部产学研结合项目(2009B090300284) |
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| 摘 要： | 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。
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| 关 键 词： | 蛇毒基因 毕赤酵母 分泌表达 |
Expression of A Snake Venom Gene with Histidine Tag in Pichia Pastoris |
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| LI Zhuo-jian.Expression of A Snake Venom Gene with Histidine Tag in Pichia Pastoris[J].Journal of Anhui Agricultural Sciences,2011,39(31):19054-19056. |
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| Authors: | LI Zhuo-jian |
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| Institution: | LI Zhuo-jian(Bioengineering Institute of Jinan University,Guangzhou,Guangdong 510632) |
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| Abstract: | 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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| Keywords: | Snake venom gene Pichia pastoris Secretory expression |
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