Team:OUC-China/Overview
From 2013.igem.org
Overview
The artificial organelle is just like an intracellular compartment which is useful in protein embellishment and that’s our origional inspiration. So we decided to reconstruct such a compartment in E.coli cells to help us transform it into cells as reactors in the future.
In previous studies,membranous organelles are the unique structure of eukaryotic cells, rare bacteria and paleontology. Magnetospirillum magneticum is considered to be an important biological model system of studying prokaryotic organelles because the structure of magnetosome in Magnetospirillum magneticum has similar traits to eukaryotic organelles with membranes. Our task is to reconstruct the magnetosome membrane in Escherichia coli, at the same time offering intracellular membranous structure (IMS) parts that can be used by prokaryotes for synthetic biology.
What also intrigued us is that the mamK gene is a gene which is crucial to IMS construction. We want to improve the mamK gene's expression by stabilizing its mRNA with a new method,hoping it can be used to promote the IMS construction. So we designed a DNA segment which we name ‘RNA guardian’, whose transcript can bind an extra ribosome. It can prevent mRNA from degradation mediated by RNase, particularly the RNase E, which has been proved to initiate the degradation of mRNA, and is usually required to interact with the 5' end for activation. So when a ribosome is binding on the mRNA near the 5' end, it can be a barrier preventing the RNaseE from interacting with the 5' end, thus preventing mRNA degradation.
Fig1.the M.magneticum AMB-1 was cultured in the magnetic spirillum growth medium(MSGM)and as you can see in the picture: the M.magneticum AMB-1 are attracted to the wall of the saline bottle and form a region like black patches.The culture medium is light red becausethe culture medium contains resazur which will turn from colorless to light red when the oxygen turn from zero to some.The bacteria tend to grown in microaerobic environment so the resazur can be regarded as an indicator for its growth.