人工sRNAs沉默<em>csrA</em>基因以优化大肠杆菌生产L-酪氨酸

中国生物工程杂志

2013, Vol. 33

Issue (8): 61-66

技术与方法

人工sRNAs沉默csrA基因以优化大肠杆菌生产L-酪氨酸

姚元锋, 赵莹, 赵广荣

天津大学化工学院制药工程系系统生物工程教育部重点实验室天津 300072

Artificial sRNAs Silencing csrA to Optimize the Production of L-tyrosine in Escherichia coli

YAO Yuan-feng, ZHAO Ying, ZHAO Guang-rong

Department of Pharmaceutical Engineering, School of Chemical Engineering and Technology, Key Laboratory of Systems Bioengineering, Ministry of Education, Tianjin University, Tianjin 300072, China

全文: PDF(1020 KB) HTML

摘要： sRNAs作为一种强有力的基因表达调控工具,在真核生物中得到了广泛的应用。随着微生物中sRNAs的不断发现以及其调控机制的逐渐明确,新近开发的人工sRNA工程在微生物代谢工程的改造上也展现了巨大的优势。通过对大肠杆菌负调控L-酪氨酸生物合成途径的碳贮藏调控因子(csrA)基因的sRNAs进行了人工设计和筛选,分析了对L-酪氨酸合成的影响。结果表明,所设计的sRNA能有效提高L-酪氨酸的合成,较高拷贝数的短anti-csrA sRNA2比较长的sRNA1 更有效,使L-酪氨酸产量提高了1.2倍。sRNA工程技术是一种有效的负调控全局代谢途径的策略,其在合成生物学以及微生物细胞工厂构建上必将有着更广泛的应用。

关键词： 人工sRNAs; 基因沉默; csrA; L-酪氨酸

Abstract: As a powerful gene regulation tool, small regulatory RNAs (sRNAs) have been widely applied in animals and plants. Recently, with the continual founding of different sRNAs and the understanding about their regulatory mechanisms in bacteria, the new developed artificial sRNAs engineering also shows a huge advantage on the microbial metabolic engineering. Carbon storage regulator (CsrA) is a post-transcriptional global regulator that has a negative role in the L-tyrosine biosynthetic pathway. By designing and screening sRNAs sequence of csrA, its effect on the L-tyrosine production in E. coli was analyzed. The results indicated that the artificial sRNAs could lead to remarkable increase of L-tyrosine production. High copy number expression of shorter anti-csrA sRNA2 which improved 1.2-fold L-tyrosine production was better than the longer sRNA1. This new strategy, being simple yet very powerful for global metabolic regulation, is thus expected to facilitate the efficient development of synthetic biology and microbial cell factories.

Key words: Artificial sRNAs Gene silencing csrA gene L-tyrosine

收稿日期: 2013-05-02 出版日期: 2013-08-25

ZTFLH:

Q819

基金资助: 国家"973"计划(2011CBA00800);国家"863"计划(2012AA02A701);天津市自然科学基金重点项目(13JCZDJC27600)资助项目

通讯作者: 赵广荣grzhao@tju.edu.cn E-mail: grzhao@tju.edu.cn

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引用本文:

姚元锋, 赵莹, 赵广荣. 人工sRNAs沉默csrA基因以优化大肠杆菌生产L-酪氨酸[J]. 中国生物工程杂志, 2013, 33(8): 61-66.

YAO Yuan-feng, ZHAO Ying, ZHAO Guang-rong. Artificial sRNAs Silencing csrA to Optimize the Production of L-tyrosine in Escherichia coli. China Biotechnology, 2013, 33(8): 61-66.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2013/V33/I8/61

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