Team:BUCT-China/Composite Part
The glyoxylate operon is a DNA fragment, and the glyoxylic acid operon designed by the team belongs to the negative control induction system of vector management. The glyoxylic acid operon is loaded into the vector plasmid and transformed into cells, which can achieve a regulatory effect, and the substrate is induced to be glyoxylic acid. When the outside is induced by glyoxylic acid, glyoxylic acid binds to the repressor protein, and the repressor protein is detached, so that transcription starts.
Source：
- Escherichia coli strain C600 chromosome, complete genome
- Sequence ID: CP031214.1
- Escherichia coli strain Es_ST2350_SE1_NDM_03_2018 chromosome, complete genome
- Sequence ID: CP031321.1
- Synthetic construct clone pEY-sRFP-A, complete sequence
- Sequence ID: KX682231.1
Design Notes
- The base sequence is edited based on the degeneracy of the protein.
- Amino acids associated with binding sites and activities cannot be altered.
- Note the addition of the necessary restriction sites
We are submitting a combination of a newly designed fatty acid operon plus a fluorescent protein that can reflect the fatty acid content with a fluorescent protein. It can be used as a molecular switch with a fatty acid signal to regulate the expression of subsequent genes. In combination with a repressor protein, RNA polymerase cannot bind to the promoter sequence, or RNA polymerase cannot move forward along the DNA, repress transcription, and then the gene cannot be expressed, and an appropriate amount of fatty acid is added to block the binding of the protein to the fatty acid. The gene can be expressed after exfoliation, and the more fatty acids, the more repressor proteins can be bound, and the better the gene expression. Using the fatty acid operon, we can add a reporter gene such as a fluorescent protein gene to the fatty acid operon to obtain a bacteria that can report the fatty acid content. The content of the fatty acid in the oil can be well reflected by the content of the fluorescent protein. An indicator of the freshness of the oil.
Source：
- Escherichia coli strain C600 chromosome, complete genome
- Sequence ID: CP031214.1
- Integration expression vector pSe1Bb1s-gfp, complete sequence
- Sequence ID: KJ814971.1
- Synthetic construct clone pEY-sRFP-A, complete sequence
- Sequence ID: KX682231.1
- Cloning vector pTHSSe_71, complete sequence
- Sequence ID: MH029788.1
Design Notes：
- Add the necessary restriction sites Pay attention to the connection and cooperation between the parts
