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author:

刘超 (刘超.) [1] | 刘树滔 (刘树滔.) [2] | 饶平凡 (饶平凡.) [3]

Abstract:

本实验旨在构建能够表达具有天然构象的重组人源铜锌超氧化物歧化酶(SOD)的毕赤酵母工程菌。通过PCR的方法从实验室保存的菌种中获得SOD目的基因后首先连接到pMD-18T载体上构建亚克隆载体,送样测序验证序列无误。对亚克隆载体使用XhoⅠ/XbaⅠ两种限制性内切酶进行酶切,获得具有粘性末端的SOD目的基因,用T4连接酶酶连到具有相同粘性末端的表达载体pPICZαA。用SacⅠ单酶切使表达载体线性化,电转化进毕赤酵母表达菌株GS115。1%甲醇诱导四天后测量酶活,筛选出酶活达530U/ml的菌株。pPICZαA载体上含有前导肽α-factor协助目的蛋白表达后分泌到酵母胞外,同时还有组氨酸标签方...

Keyword:

毕赤酵母 表达 超氧化物歧化酶(SOD)

Community:

  • [ 1 ] 福州大学生物工程研究所

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Year: 2012

Language: Chinese

Cited Count:

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ESI Highly Cited Papers on the List: 0 Unfold All

WanFang Cited Count:

Chinese Cited Count:

30 Days PV: 1

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