Optogenetic regulation of artificial microRNA improves H(2) production in green alga Chlamydomonas reinhardtii.
Optogenetic regulation of artificial microRNA improves H(2) production in green alga Chlamydomonas reinhardtii.
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人工 microRNA 的光遗传学调控可提高绿藻莱茵衣藻的 H-2 产量
DOI：
10.1186/s13068-017-0941-7
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发表时间：
2017
影响因子：
6.3
通讯作者：
Hu Z
中科院分区：
文献类型：
--
作者：
Wang Y;Jiang X;Hu C;Sun T;Zeng Z;Cai X;Li H;Hu Z
莱茵衣藻是一种理想的模式生物，不仅可用于研究植物和动物的基本代谢过程，还可用于生产包括氢气在内的生物燃料。莱茵衣藻的转基因分析现已成熟且非常方便，但诱导型外源基因表达系统仍有待研究。最常用的热激诱导系统对藻类细胞生长有严重影响，并且在大规模培养中控制起来困难且成本高昂。先前对衣藻光产氢的研究也利用这种热诱导系统来激活靶基因转录和氢合成。在这里，我们描述了一种蓝光诱导系统，通过该系统我们实现了莱茵衣藻目标基因表达的光遗传学调控。这种光诱导系统首次在光合生物中被设计。光诱导异二聚化蛋白 CRY2 和 CIB1 分别与 VP16 转录激活结构域和 GAL4 DNA 结合结构域融合。该方案允许响应蓝光而转录激活激活序列下游的靶基因。利用该系统，我们成功地改造了蓝光诱导产氢转基因藻类。转基因藻在红光下培养并大致正常生长直至对数生长期。当蓝光照射时，转基因藻类表达针对光合系统 D1 蛋白的人工 miRNA，并观察到氢气产量的改变。光诱导系统成功激活了人工 miRNA，从而在蓝光下对其靶基因进行了调控。此外，使用该系统提高了产氢量，这表明在大规模微藻培养中基因表达调控提供了一种更方便、更有效的方法。这种光遗传学基因控制系统是基因调控的有用工具，也建立了一种提高绿藻产氢的新方法。本文的在线版本 (10.1186/s13068-017-0941-7) 包含补充材料，可供授权用户使用。
Chlamydomonas reinhardtii is an ideal model organism not only for the study of basic metabolic processes in both plants and animals but also the production of biofuels including hydrogen. Transgenic analysis of C. reinhardtii is now well established and very convenient, but inducible exogenous gene expression systems remain under-studied. The most commonly used heat shock-inducible system has serious effects on algal cell growth and is difficult and costly to control in large-scale culture. Previous studies of hydrogen photoproduction in Chlamydomonas also use this heat-inducible system to activate target gene transcription and hydrogen synthesis. Here we describe a blue light-inducible system with which we achieved optogenetic regulation of target gene expression in C. reinhardtii. This light-inducible system was engineered in a photosynthetic organism for the first time. The photo-inducible heterodimerizing proteins CRY2 and CIB1 were fused to VP16 transcription activation domain and the GAL4 DNA-binding domain, respectively. This scheme allows for transcription activation of the target gene downstream of the activation sequence in response to blue light. Using this system, we successfully engineered blue light-inducible hydrogen-producing transgenic alga. The transgenic alga was cultured under red light and grew approximately normally until logarithmic phase. When illuminated with blue light, the transgenic alga expressed the artificial miRNA targeting photosynthetic system D1 protein, and altered hydrogen production was observed. The light-inducible system successfully activated the artificial miRNA and, consequently, regulation of its target gene under blue light. Moreover, hydrogen production was enhanced using this system, indicating a more convenient and efficient approach for gene expression regulation in large-scale microalgae cultivation. This optogenetic gene control system is a useful tool for gene regulation and also establishes a novel way to improve hydrogen production in green algae. The online version of this article (10.1186/s13068-017-0941-7) contains supplementary material, which is available to authorized users.
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影响因子：
4.6
作者：
Nakajima M;Ferri S;Rögner M;Sode K
通讯作者：
Sode K
影响因子：
10.5
作者：
Zhao, Tao;Li, Guanglin;Qi, Yijun
通讯作者：
Qi, Yijun
影响因子：
7.4
作者：
Melis, A;Zhang, LP;Seibert, M
通讯作者：
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影响因子：
56.9
作者：
Liu, Hongtao;Yu, Xuhong;Lin, Chentao
通讯作者：
Lin, Chentao
DOI：
10.1016/j.bbamcr.2010.12.004
发表时间：
2011-02-01
影响因子：
5.1
作者：
Bodvard, Kristofer;Wrangborg, David;Kall, Mikael
通讯作者：
Kall, Mikael