Difference between revisions of "Part:BBa K4863003"
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Slp is a native surface exposed protein that constitutes of the S-layer on the outermost surface of <i>Synechocystis sp.</i>. Fusion of a recombinant protein to Slp will yield a cell completely covered with the fusion protein on the outermost surface. SpyTag is suitable for fusion with Slp since fusion of larger proteins with Slp will likely impede growth of the cell as we have learned. Via the displaying SpyTag, a protein complex of SpyCatcher fused with a large protein can be displayed on the cell surface of engineered <i>Synechocystis</i> via covalent bonding between SpyTag and SpyCatcher. | Slp is a native surface exposed protein that constitutes of the S-layer on the outermost surface of <i>Synechocystis sp.</i>. Fusion of a recombinant protein to Slp will yield a cell completely covered with the fusion protein on the outermost surface. SpyTag is suitable for fusion with Slp since fusion of larger proteins with Slp will likely impede growth of the cell as we have learned. Via the displaying SpyTag, a protein complex of SpyCatcher fused with a large protein can be displayed on the cell surface of engineered <i>Synechocystis</i> via covalent bonding between SpyTag and SpyCatcher. | ||
| − | SpyTag is a 13 amino acid-long peptide chain originally isolated from <i>Streptococcus pyogenes</i>. Slp is encoded by the gene sll1951 in Synechocystis PCC6803. This protein forms a lattice that is about 30nm wide that covers the cell surface, forming the paracrystalline S-layer. It has an undefined signal peptide at C-terminus. | + | SpyTag is a 13 amino acid-long peptide chain originally isolated from <i>Streptococcus pyogenes</i>. Slp is encoded by the gene sll1951 in <i>Synechocystis</i> PCC6803. This protein forms a lattice that is about 30nm wide that covers the cell surface, forming the paracrystalline S-layer. It has an undefined signal peptide at C-terminus. |
===Characterization=== | ===Characterization=== | ||
Revision as of 04:02, 11 October 2023
PpsbA2_SpyTag_Slp
A SpyTag-Slp protein complex driven by a light-inducible promoter. SpyTag (BBa_K1159200) binds to the N-terminus of surface layer protein (Slp:BBa_K4863002) of Synechocystis PCC 6803 to be displayed on the cell surface of the engineered organism.
Usage and Biology
Slp is a native surface exposed protein that constitutes of the S-layer on the outermost surface of Synechocystis sp.. Fusion of a recombinant protein to Slp will yield a cell completely covered with the fusion protein on the outermost surface. SpyTag is suitable for fusion with Slp since fusion of larger proteins with Slp will likely impede growth of the cell as we have learned. Via the displaying SpyTag, a protein complex of SpyCatcher fused with a large protein can be displayed on the cell surface of engineered Synechocystis via covalent bonding between SpyTag and SpyCatcher.
SpyTag is a 13 amino acid-long peptide chain originally isolated from Streptococcus pyogenes. Slp is encoded by the gene sll1951 in Synechocystis PCC6803. This protein forms a lattice that is about 30nm wide that covers the cell surface, forming the paracrystalline S-layer. It has an undefined signal peptide at C-terminus.
Characterization
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 3552
Illegal EcoRI site found at 5532
Illegal XbaI site found at 758
Illegal PstI site found at 874
Illegal PstI site found at 4474 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 3552
Illegal EcoRI site found at 5532
Illegal NheI site found at 1181
Illegal NheI site found at 1278
Illegal PstI site found at 874
Illegal PstI site found at 4474 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 3552
Illegal EcoRI site found at 5532
Illegal BglII site found at 2693
Illegal BamHI site found at 3599
Illegal BamHI site found at 4913
Illegal XhoI site found at 816 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 3552
Illegal EcoRI site found at 5532
Illegal XbaI site found at 758
Illegal PstI site found at 874
Illegal PstI site found at 4474 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 3552
Illegal EcoRI site found at 5532
Illegal XbaI site found at 758
Illegal PstI site found at 874
Illegal PstI site found at 4474
Illegal AgeI site found at 2580
Illegal AgeI site found at 2763
Illegal AgeI site found at 3111
Illegal AgeI site found at 3171
Illegal AgeI site found at 4692 - 1000COMPATIBLE WITH RFC[1000]