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家蝇抗真菌肽MAF-1原核表达条件优化及活性验证

DOI: 10.11847/zgggws2015-31-11-16, PP. 1420-1423

Keywords: 家蝇抗真菌肽(MAF-1),重组融合蛋白,表达条件,纯化

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Abstract:

?目的优化家蝇抗真菌肽(MAF-1)原核表达条件及重组蛋白的活性验证。方法利用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和QuantityOne凝胶电泳图像分析系统研究不同的诱导温度、异丙基硫代-β-D半乳糖苷(IPTG)诱导浓度和诱导时间对融合蛋白表达的影响;融合蛋白用镍离子金属螯合剂亲和层析柱进行蛋白纯化,切除His标签后进行活性验证。结果在加入浓度为25μmol/L的IPTG,34℃诱导18h,融合蛋白的表达量最高;融合蛋白纯化后浓度为0.706mg/mL,其最低抑杀白色念珠菌的浓度为70μg/mL。结论成功获得重组融合蛋白的最佳表达条件,并纯化目的蛋白,切除His标签后的目的蛋白具有抗真菌活性。

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