Difference between revisions of "Part:BBa K5136035"

Revision as of 02:37, 28 September 2024

inpnc-His tag-SpyCatcher

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NotI site found at 481
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 330
Illegal XhoI site found at 1089
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 72
Illegal NgoMIV site found at 405
Illegal AgeI site found at 429
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K5136035 short</partinfo>
 <!-- Add more about the biology of this part here
 ===Usage and Biology===
+Biology
+INPNC
+Ice nucleoprotein (INP), an outer membrane protein from Pseudomonas syringae, has been used as a surface anchor in many researches. The truncated version of INP, namely INPNC which contains only the N- and C-terminal portion of INP, has excellent capacity to anchor target proteins on the cell membrane (1).
+SpyCatcher
+SpyCatcher is a engineered split fragment of the fibronectin-binding protein (FbaB) in Streptococcus pyogenes, containing 116 residues. SpyTag is a peptide tag of 13 residues developed by previous researchers. SpyCatcher could steadily bind to SpyTag by forming an isopeptide bond (2).
+Usage
+We aim to displaying metallothioneins MT2A and MT3 on the surface of E. coli by INPNC, which could improve the adsorption rate of heavy metal ions for the harmless treatment of deinking wastewater. However, it is tricky to verify whether a protein has been successfully displayed on the surface of bacteria. So, we introduce the Spy system. In our project, SpyCatcher is fused with INPNC and SpyTag is fused with GFP. Then, engineered bacteria which express the fused protein INPNC-his tag-SpyCatcher are treated with SpyTag-GFP. By monitoring the fluorescence intensity of bacteria, we can verify if INPNC could anchor target proteins on cell surface. Here, we constructed the composite part (BBa_K5136224) to express INPNC-his tag-SpyCatcher.
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 <span class='h3bb'>Sequence and Features</span>