Team:Saint Joseph/Parts Overview
Parts Overview
How SHuffle T7 E.coli strain works?
Chemically competent E. coli K12 cells suitable for T7 protein expression with enhanced capacity to correctly fold proteins with multiple disulfide bonds in the cytoplasm. .
Key Advantages
- 1. Transformation efficiency: 1 x 106 cfu/µg pUC19 DNA
- 2. Engineered E. coli K12 to promote disulfide bond formation in the cytoplasm
- 3. Constitutively expresses a chromosomal copy of the disulfide bond isomerase DsbC
- 4. DsbC promotes the correction of mis-oxidized proteins into their correct form (1,3)
- 5. The cytoplasmic DsbC is also a chaperone that can assist in the folding of proteins that do not require disulfide bonds (4)
- 6. Expresses a chromosomal copy of T7 RNAP
- 7. Tight control of expression by lacIq allows potentially toxic genes to be cloned
- 8. Resistance to phage T1 (fhuA2), Nit, Str, Spec.
How PET26b works?
The pET-26b(+) vector carries an N-terminal pelB signal sequence for potential periplasmic localization, plus optional C-terminal His•Tag® sequence. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the vector map (TB071)
What is Pelb leader sequence?
The pelb leader sequence is a sequence of amino acids which, when attached to a protein, directs the protein to the bacterial periplasm, where the sequence is removed by a signal peptidase. The leader sequence consists of the 22 N-terminal amino acid residues.
How It works?
This leader sequence can be attached to any other protein (on the DNA level) resulting in a transfer of such a fused protein to the periplasmic space of gram negative bacteria, such as Escherichia Coli, often used in genetic engineering.
Saint Joseph BioBrick Parts
| Name | Type | Description | Designer | Length |
|---|---|---|---|---|
| BBa_K2962000 | Coding | lcc1 gene codes laccase enzyme | Handan Kesim | 1575 |
| BBa_K2962001 | Plasmid_Backbone | PET26b | Handan Kesim | 5360 |
| BBa_K2962002 | Tag | jdsb fusion tag | Handan Kesim | 17 |
| BBa_K2962003 | Tag | histidine tag | Handan Kesim | 18 |
| BBa_K2962004 | Signalling | Signalling | Handan Kesim | 63 |
| BBa_K2962005 | Cell | SHuffle T7 E.coli strain | Handan Kesim | - |
| BBa_K2962006 | Composite | pET26b with laccase enzyme from Trametes Versicolor. | Sera Küçük | 6943 |
| BBa_K2962007 | Composite | pET26b with pelB laccase (signal peptide laccase) | Sera Küçük | 5431 |