- 著者
- Satoru Watanabe Shunsuke Saito Yasuhiro Suezaki Takeshi Seguchi Ryudo Ohbayashi
- 出版者
- Applied Microbiology, Molecular and Cellular Biosciences Research Foundation
- 雑誌
- The Journal of General and Applied Microbiology (ISSN:00221260)
- 巻号頁・発行日
- pp.2019.11.005, (Released:2020-02-25)
- 参考文献数
- 18
- 被引用文献数
- 1
In bacterial DNA replication, the initiator protein DnaA binds to the multiple DnaA box sequences located at oriC to facilitate the unwinding of duplex DNA strands. The cyanobacterium Synechococcus elongatus PCC 7942, which contains multiple chromosomal copies per cell, has DnaA box (like sequences around the oriC region, which is located upstream of dnaN. We previously observed the binding of DnaA around the oriC region; however, the DNA-binding specificity of DnaA to DnaA box sequences has not been examined. Here, we analyzed the binding specificity of DnaA protein to the DnaA box in S. elongatus by using bio-layer interferometry (BLI), a method for monitoring intermolecular interactions. We observed that recombinant DnaA protein recognized specifically the DnaA box sequence TTTTCCACA in vitro. In addition, DNA binding activity was significantly increased by R328H mutation of DnaA. This is the first report to characterize DnaA binding to the DnaA box sequence in cyanobacteria.
- 著者
- Ryudo Ohbayashi Hideto Akai Hirofumi Yoshikawa Wolfgang R. Hess Satoru Watanabe
- 出版者
- 公益財団法人 応用微生物学・分子細胞生物学研究奨励会
- 雑誌
- The Journal of General and Applied Microbiology (ISSN:00221260)
- 巻号頁・発行日
- pp.2016.02.002, (Released:2016-06-01)
- 参考文献数
- 32
- 被引用文献数
- 43
Cyanobacteria are photosynthetic microorganisms that serve as experimental model organisms for the study of photosynthesis, environmental stress responses, and the production of biofuels. Genetic tools for bioengineering have been developed as a result of such studies. However, there is still room for improvement for the tight control of experimental protein expression in these microorganisms. Here, we describe an expression system controlled by a theophylline-responsive riboswitch that we have constructed in the cyanobacterium Synechocystis sp. PCC 6803. We demonstrate that, in response to different theophylline concentrations, this riboswitch can tightly control green fluorescence protein expression in Synechocystis. Thus, this system is useful as a tool for genetic engineering and the synthetic biology of cyanobacteria.