Autor: |
Weber N; Botanical Institute, Karlsruhe Institute of Technology KIT., Hofmeister M; Botanical Institute, Karlsruhe Institute of Technology KIT., Wunsch N; Botanical Institute, Karlsruhe Institute of Technology KIT., Kohler A; Botanical Institute, Karlsruhe Institute of Technology KIT., Kaster AK; Institute for Biological Interfaces 5 (IBG 5), Karlsruhe Institute of Technology KIT., Vollmers J; Institute for Biological Interfaces 5 (IBG 5), Karlsruhe Institute of Technology KIT., Kachel B; Institut für Technische Mikrobiologie, Hochschule Mannheim., Mack M; Institut für Technische Mikrobiologie, Hochschule Mannheim., Lamparter T; Botanical Institute, Karlsruhe Institute of Technology KIT; tilman.lamparter@kit.edu. |
Abstrakt: |
Cyanobacteria are the focus of basic research and biotechnological projects in which solar energy is utilized for biomass production. Phormidium lacuna is a newly isolated filamentous cyanobacterium. This paper describes how new filamentous cyanobacteria can be isolated from marine rockpools. It also describes how DNA can be extracted from filaments and how the genomes can be sequenced. Although transformation is established for many single-celled species, it is less frequently reported for filamentous cyanobacteria. A simplified method for the natural transformation of P. lacuna is described here. P. lacuna is the only member of the order Oscillatoriales for which natural transformation is established. This paper also shows how natural transformation is used to express superfolder green fluorescent protein (sfGFP). An endogenous cpcB promoter induced approximately 5 times stronger expression than cpc560, A2813, or psbA2 promoters from Synechocystis sp. PCC6803. Further, a method for the cryopreservation of P. lacuna and Synechocystis sp. CPP 6803 was established, and methods for assessing motility in a liquid medium and on agar and plastic surfaces are described. |