Part:BBa_K1727002
Signal peptide (S.lividans chitinase C)
Mechanism of signal peptide
This signal peptide is comes from the signal of chitinase C that extracted by S.lividans. It had been proved that can worked in E.coli and successfully secret target protein to LB culture medium. After translation process signal peptide will lead mature peptide to the secretion system of E.coli. When the pre-mature peptides go through the periplasmic space, peptidase will identified the cleavage site Ala-Gln-Ala and cut at the double Ala at the signal and mature peptide linkage site. Then separate signal peptide from mature peptide. Finally, secreting out mature peptide into the culture medium.
Features of signal peptide
Small and simple: The sequence of Signiferin is only around 100 bp. For E.coli, 100 base pairs is in the working capacity.
This part is a coding gene from S.lividans chitinase C.
|
|
Fig.1 Gel electrophoresis for biobrick BBa_K1727002 Finally, we used EcoR I and Pst I these two kinds of enzyme to check the insert really contain in the biobrick. We used 2% gel. The size of signal peptide insert should be 100 to 200 bp. There are bands on all the sample of enzyme digest product. As the result, we can make sure that the signal peptide has been ligated in backbone. |
Experimentally validate
Fig.2 Chart of Signiferin disinfection ability to S.aureus
We had tested the function of biobrick BBa_K1727000 it built as the fusion peptide of sinal peptide and Signiferin. After IPTG induction process we picked LB medium used centrifuge and filter to remove the bacteria than test the ability of Signiferin. As the chart we can see that Signifein has the functions to kill S.aureus. So we can ensure signal peptide is work in our system.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
| None |