| Non-Scientific Abstract Bacteria have many different methods of storing
carbon. Some bacteria store carbon as granules of polyhydroxyalkanoic acids (PHAs). PHAs are biodegradable
plastics, and it is believed that they have great potential as biomaterials. The goal of this research is
to understand the regulation of the synthesis of bioplastics.Once the mechanisms are understood it is
possible to capitalize on the production of this environmentally friendly material instead of conventional
plastics. In this study, a protein, PhaR, was investigated to determine its role in polyhydroxyalkanoic
acid synthesis. It was determined that PhaR bound to a section of DNA. Specifically, PhaR bound to a
region of the phaP gene. Through laboratory experimentation, it was found that the interaction
between the PhaR protein and the phaP gene was specific and can be further investigated to
determine its role in regulation of carbon storage.As the understanding of this system increases, the use
of biodegradeable plastics made from bacteria could become an important advancement in commercially used
materials.
Abstract
Polyhydroxyalkanoic acids (PHAs) are produced in many bacteria and accumulated as intracellular
granules. PhaP (phasin) is a protein produced by the phaP gene that binds to PHA granules and promotes
further PHA synthesis. phaP is suspected to be regulated by PhaR.This transcriptional regulatory protein,
PhaR was investigated in this study. This protein is believed to be involved in polyhydroxyalkanoic acid
synthesis by negatively regulating phaP. A gel mobility shift assay, using Streptavidin-Biotin detection,
showed that PhaR specifically bound to the promoter region in a segment located between 105 and 312 base
pairs upstream of the start codon of the phaP gene. These results suggest that PhaR is a DNA-binding
protein that may play a role in the regulation of phaP gene expression. The phaR and the phaP promoter
segment were transferred from Ralstonia eutropha into Escherichia coli and investigated in that
manner. |
