Design and characterization of molecular tools for a Synthetic Biology approach towards developing cyanobacterial biotechnology.

Design and characterization of molecular tools for a Synthetic Biology approach towards developing cyanobacterial biotechnology.
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DOI:
10.1093/nar/gkq164
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发表时间:
2010-05
影响因子:
14.9
通讯作者:
Heidorn T
Heidorn T
中科院分区:
生物学2区
文献类型:
--
作者:
Huang HH;Camsund D;Lindblad P;Heidorn T

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蓝细菌适用于可持续的太阳能生物技术应用。合成生物学将生物学与计算设计和工程学观点联系起来,但需要有效的工具和有关生物部件和系统功能的信息。为了使蓝藻合成生物学的发展,开发和表征了几种分子工具:(i)构建了广泛宿主范围的BioBrick穿梭载体pPMQAK 1,并证实其在大肠杆菌和三种不同的蓝藻菌株中复制。(ii)荧光蛋白Cerulean,GFPmut3B和EYFP已被证明是蓝藻中的报告蛋白,尽管光合色素的强背景。(iii)几个启动子,如PrnpB和PrbcL的变体,和一个版本的启动子Ptrc与两个运营商增强镇压,开发和集胞藻属菌株PCC 6803的特点。(iv)结果表明,在集胞藻属菌株PCC 6803中,能够进行动态表达研究所需的靶向蛋白质降解系统正在发挥作用。pPMQAK1穿梭载体允许在许多原核生物中使用越来越多的BioBrick部分,并且本文实施的其他工具促进了蓝细菌中新部分和系统的开发。
Cyanobacteria are suitable for sustainable, solar-powered biotechnological applications. Synthetic biology connects biology with computational design and an engineering perspective, but requires efficient tools and information about the function of biological parts and systems. To enable the development of cyanobacterial Synthetic Biology, several molecular tools were developed and characterized: (i) a broad-host-range BioBrick shuttle vector, pPMQAK1, was constructed and confirmed to replicate in Escherichia coli and three different cyanobacterial strains. (ii) The fluorescent proteins Cerulean, GFPmut3B and EYFP have been demonstrated to work as reporter proteins in cyanobacteria, in spite of the strong background of photosynthetic pigments. (iii) Several promoters, like PrnpB and variants of PrbcL, and a version of the promoter Ptrc with two operators for enhanced repression, were developed and characterized in Synechocystis sp. strain PCC6803. (iv) It was shown that a system for targeted protein degradation, which is needed to enable dynamic expression studies, is working in Synechocystis sp. strain PCC6803. The pPMQAK1 shuttle vector allows the use of the growing numbers of BioBrick parts in many prokaryotes, and the other tools herein implemented facilitate the development of new parts and systems in cyanobacteria.
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