Difference between revisions of "Part:BBa K3098009"

Revision as of 17:01, 21 October 2019

GST+PgD5+Avitag

This part is the one we are hoping to use in the production of the functional PgD5.

usage and biology

In this part, GST tag helps the dissolution and purification of the peptide while Avitag provides a site for biotinylation as a linkage to the dCBM3a and BC. It has been proved by our experiments that these modifications on the peptide will not eliminate the antifungal function of the peptides after our corresponding treatment, so we'll be able to produce functional antimicrobial peptides with this precursor.

WHU-China 2019

Expression quantity of PgD5-Avitag is so low that it could not be purified well. To enhance the expression quantity,we add GST-Tag at N-terminal, it could increase the solubility of fusion protein and avoid the inclusion body. We induced the expression of GST-PgD5 fusion protein and purified it by GST affinity column and eluted the fusion protein by GSH. We did SDS-PAGE electrophoresis, the below was the result.

The GST-Tag can be cleaved by PreScission Protease. Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 688
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 784
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI.rc site found at 85

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K3098009 short</partinfo>
-This part is the one we are hoping to use in the production of the functional PgD5. In this part, GST tag helps the dissolution and purification of the peptide while Avitag provides a site for biotinylation as a linkage to the dCBM3a and BC. It has been proved by our experiments that these modifications on the peptide will not eliminate the antifungal function of the peptides after our corresponding treatment, so we'll be able to produce functional antimicrobial peptides with this precursor.
+This part is the one we are hoping to use in the production of the functional PgD5.
 ===usage and biology===
+In this part, GST tag helps the dissolution and purification of the peptide while Avitag provides a site for biotinylation as a linkage to the dCBM3a and BC. It has been proved by our experiments that these modifications on the peptide will not eliminate the antifungal function of the peptides after our corresponding treatment, so we'll be able to produce functional antimicrobial peptides with this precursor.
 ===WHU-China 2019===
 Expression quantity of PgD5-Avitag is so low that it could not be purified well. To enhance the expression quantity,we add GST-Tag at N-terminal, it could increase the solubility of fusion protein and avoid the inclusion body. We induced the expression of GST-PgD5 fusion protein and purified it by GST affinity column and eluted the fusion protein by GSH. We did SDS-PAGE electrophoresis, the below was the result.