Nozzi Nicole E, Atsumi Shota
Department of Chemistry, University of California, Davis , One Shields Avenue, Davis, California 95616, United States.
ACS Synth Biol. 2015 Nov 20;4(11):1197-204. doi: 10.1021/acssynbio.5b00057. Epub 2015 May 19.
Cyanobacteria have gained popularity among the metabolic engineering community as a tractable photosynthetic host for renewable chemical production. However, though a number of successfully engineered production systems have been reported, long-term genetic stability remains an issue for cyanobacterial systems. The genetic engineering toolbox for cyanobacteria is largely lacking inducible systems for expression control. The characterization of tight regulation systems for use in cyanobacteria may help to alleviate this problem. In this work we explore the function of the IPTG inducible promoter P(L)lacO1 in the model cyanobacterium Synechococcus elongatus PCC 7942 as well as the effect of gene order within an operon on pathway expression. According to our experiments, P(L)lacO1 functions well as an inducible promoter in S. elongatus. Additionally, we found that gene order within an operon can strongly influence control of expression of each gene.
蓝细菌作为用于可再生化学品生产的易于操作的光合宿主，在代谢工程领域受到了广泛关注。然而，尽管已经报道了许多成功构建的生产系统，但长期遗传稳定性仍然是蓝细菌系统面临的一个问题。蓝细菌的基因工程工具箱在很大程度上缺乏用于表达控制的诱导系统。用于蓝细菌的严格调控系统的表征可能有助于缓解这一问题。在这项工作中，我们探索了IPTG诱导型启动子P(L)lacO1在模式蓝细菌聚球藻PCC 7942中的功能，以及操纵子内基因顺序对途径表达的影响。根据我们的实验，P(L)lacO1在聚球藻中作为诱导型启动子功能良好。此外，我们发现操纵子内的基因顺序可以强烈影响每个基因的表达控制。
