| The concept of introducing engineering principles of abstraction and standardization into synthetic biology has received increasing attention in the past several years and continues to be in the forefront of synthetic biology. One direction being pursued by synthetic biologists is creation of modular biological parts (BioBrick(TM)) that can be readily synthesized and mixed together in different combinations. However, most standard BioBrick(TM) parts in the Registry were designed for E. coli, although synthesis of specific BioBrick(TM) parts for other bacteria, such as for yeast and cyanobacteria, have begun. Besides, at the present time, there are only three chassis, which include E. coli, Bacillus subtilis, and a cell-free chassis, available in the Registry. Thus, the choices of BioBrick(TM) chassis are very limited. In addition, most BioBrick(TM) parts in the Registry have not been characterized.;In the present study, the BioBrick(TM) concept was extended to the photosynthetic bacteria, Rhodobacter sphaeroides. In order to do that, a BioBrick(TM) compatible gene expression system was designed to convert R. sphaeroides to potential solar powered bio-factories or bio-refineries. This gene expression system was composed of BioBrick(TM) promoters, Ribosome Binding Sites (RBSs), and terminators in a BioBrick(TM) compatible cloning vector and its function has been validated through the expression of fluorescent proteins.;In addition, a bioluminescence-based BioBrick(TM) characterization method was developed in this study. This method was based on a cloning vector that includes two adjacent operons, with each expressing a different luciferase reporter gene. The measured optical signals from the two expressed bioluminescent reporters were then used to predict the performance of promoters, RBSs, and terminators.;Based on this bioluminescence-based BioBrick(TM) characterization method, two BioBrick(TM) characterizations kits, one for E. coli and one for R. sphaeroides, were developed. BioBrick(TM) parts that include seven promoters, six RBSs, and six terminators were characterized using the E. coli characterization kit. R. sphaeroides BioBrick(TM) parts were characterized when R. sphaeroides containing the BioBrick(TM) measurement constructs were cultured by both anaerobic photosynthesis and by aerobic respiration respectively. The experimental results showed that the activities of these R. sphaeroides BioBrick(TM) parts were very similar for the cells growing under two different conditions. |
