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Team:Tokyo-NoKoGen/Project/aggregation
From 2010.igem.org
Aggregation
Introduction
In our EcoTanker, the goal is to collect objective substance automatically by using E. coli. However, to collect substance, we have to collect the cell at first. So we aimed to construct a device, which signals E. coli to self-aggregate and we focused on a protein, Antigen 43. This protein is derived from E. coli.
Antigen 43 consists of two protein subunits, α and β, with apparent molecular masses of about 50 and 53 kDa. The β subunit is attached to the cell surface via interaction with the β subunit, which is an integral outer membrane component. Antigen 43 has the N-terminal signal peptide and it directs translocation across the cytoplasmic membrane to the periplasm via the general secretory pathway. Subsequently, the β domain forms a β barrel structure in the outer membrane through which the β domain gains access to the cell exterior [1].
Antigen 43 consists of two protein subunits, α and β, with apparent molecular masses of about 50 and 53 kDa. The β subunit is attached to the cell surface via interaction with the β subunit, which is an integral outer membrane component. Antigen 43 has the N-terminal signal peptide and it directs translocation across the cytoplasmic membrane to the periplasm via the general secretory pathway. Subsequently, the β domain forms a β barrel structure in the outer membrane through which the β domain gains access to the cell exterior [1].
Why is this device needed?
Aggregation device is needed because
- Taking up a target material in E. coli.
- Self-aggregation of E. coli. : Skipping the harvest work.
What is this device composed inside it?
This device expresses Antigen 43, which is encoded by the gene agn43. For the promoter, we used OmpR (+) promoter (ompC promoter) (BBa_R0082)(Fig. 1). This promoter responds to phosphorylated OmpR (response regulator). OmpR will phosphorylated by EnvZ, which is a histidine kinase.
