草菇过氧化氢酶基因(VvCAT1)异源表达及耐温度胁迫功能分析

Expression of catalase gene (VvCAT1) from Volvariella volvacea in Escherichia coli and its temperature tolerance

摘要:

[背景] 过氧化氢酶（catalase，CAT）参与真菌的生长发育，逆境胁迫时保护真菌免受氧化损伤。[目的] 实现草菇过氧化氢酶基因（VvCAT1）的异源表达，分析VvCAT1耐温度胁迫的功能。[方法] 克隆VvCAT1，构建过表达载体pBAR GPE1/VvCAT1，转化到大肠杆菌（Escherichia coli）菌株Stbl3中，异源表达草菇过氧化氢酶。测定温度胁迫后重组菌（pBAR GPE1/VvCAT1/Stbl3）与对照菌（pBAR GPE1/Stbl3）的过氧化氢酶活性和生长情况，验证VvCAT1的功能。[结果] 重组菌的CAT酶活性显著提高，生长情况显著优于对照菌。[结论] VvCAT1的导入及表达显著提高了大肠杆菌Stbl3的耐温度胁迫功能。

Abstract:

[Background] Catalase (CAT) is involved in the growth and development of fungi and protects them from oxidative damage in the event of stress. [Objective] The VvCAT1 gene from Volvariella volvacea was expressed in Escherichia coli Stbl3 and its temperature tolerance was characterized. [Methods] VvCAT1 was cloned to construct a recombinant plasmid pBAR GPE1/VvCAT1 which was then transformed into Stbl3 to yield a recombinant strain expressing catalase. Catalase activity and growth of the recombinant strain (pBAR GPE1/VvCAT1/Stbl3) and the control strain (pBAR GPE1/Stbl3) were measured to verify the temperature tolerance. [Results] The recombinant strain exhibited stronger heat and cold tolerance than the control with increased catalase activity and better growth. [Conclusion] The transgenosis and expression of VvCAT1 significantly improved the heat and cold tolerance of E. coli Stbl3.