大肠杆菌menA敲除对CoQ积累的影响研究

doi:10.13560/j.cnki.biotech.bull.1985.2015.12.030

摘要/Abstract

摘要：

通过同源重组敲除大肠杆菌的menA基因，增加菌体CoQ合成量，用于构建CoQ高产菌株。以pKD4质粒为模板，PCR扩增kan^r片段；在pKD46的辅助下，kan^r片段转化大肠杆菌，利用抗生素筛选和PCR验证重组子；以紫外诱变菌为对照，发酵menA基因敲除菌株，分析CoQ种类和产量变化。成功获得menA基因敲除菌株，CoQ种类不变，产量增加约为38%。首次敲除menA基因，改良株CoQ产量得到提高，达到预期目标。

关键词: 大肠杆菌, 基因敲除, menA基因, CoQ, 同源重组

Abstract:

Knocking out menA gene of Escherichia coli by homologous recombination increases the synthetized amount of CoQ, which is used for constructing the strains of high-yield CoQ. Using pKD4 plasmid as template, the kan^r fragments were amplified by PCR. In the presence of auxiliary plasmid pKD46, the kan^r fragments were transformed into Escherichia coli, and the recombinant one was confirmed by antibiotic screening and verification of PCR；By contrast with uv-induced mutation, the strains with menA gene knocked out were fermented, and the types of CoQ and the changes of CoQ yield were analyzed. The results showed that the strains with menA gene knockout were successfully obtained, while the types of CoQ unchanged and the yield increased about 38%. In conclusion, this is the first time of knocking out menA gene, CoQ production of mutant strains is improved, and the desired goal is achieved, which lays a foundation for the further construction of high-yield CoQ strains.

Key words: Escherichia coli, gene knockout, menA genes, CoQ, homologous recombination

刘永青,任琳,张子锋. 大肠杆菌menA敲除对CoQ积累的影响研究[J]. 生物技术通报, 2015, 31(12): 207-213.

Liu Yongqing, Ren Lin, Zhang Zifeng. The Influence of Knockout of menA Gene in Escherichia coli on the Accumulation of CoQ[J]. Biotechnology Bulletin, 2015, 31(12): 207-213.

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