Difference between revisions of "Part:BBa K3739041"
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LMT<br/>Lytic murein transglycosylase (LMT) is an enzyme which is able to degrade murein, a component of cell wall of bacteria. There are two kind of LMTs existing in ''E.coli'': the membrane-binding one and the soluble one. The gene coding LMT homolog is also incorporated in the genome of ''Vibrio natriegens''. The LMT signal peptide (named LMT in our parts) is from the LMT homolog, which can lead the fused protein secreted out of ''Vibrio natriegens''.<br/>GFP<br/>Protein tags are peptides genetically fused to the proteins of interest. There are many different kinds of tags developed for for different purposes. For labeling experiments, normally the proteins of interest are tagged directly by fluorescent tags. Green fluorescent protein (GFP) make it easier to study protein localization, interactions and dynamics when fusing with other peptides and proteins. | LMT<br/>Lytic murein transglycosylase (LMT) is an enzyme which is able to degrade murein, a component of cell wall of bacteria. There are two kind of LMTs existing in ''E.coli'': the membrane-binding one and the soluble one. The gene coding LMT homolog is also incorporated in the genome of ''Vibrio natriegens''. The LMT signal peptide (named LMT in our parts) is from the LMT homolog, which can lead the fused protein secreted out of ''Vibrio natriegens''.<br/>GFP<br/>Protein tags are peptides genetically fused to the proteins of interest. There are many different kinds of tags developed for for different purposes. For labeling experiments, normally the proteins of interest are tagged directly by fluorescent tags. Green fluorescent protein (GFP) make it easier to study protein localization, interactions and dynamics when fusing with other peptides and proteins. | ||
===Usage=== | ===Usage=== | ||
| − | Here, we used BBa_K3739010 to construct the expression system and obtained the composite part BBa_K3739041. We transformed the constructed plasmid on the expression vector pET-28a (+) into V. natriegens to extract the protein. This protein work together with those from BBa_K3739038 and BBa_K3739043 to verify the function of our secretory peptides Aly01 and LMT. | + | Here, we used BBa_K3739010 to construct the expression system and obtained the composite part BBa_K3739041. We transformed the constructed plasmid on the expression vector pET-28a (+) into ''V. natriegens'' to extract the protein. This protein work together with those from BBa_K3739038 and BBa_K3739043 to verify the function of our secretory peptides Aly01 and LMT. |
===Characterization=== | ===Characterization=== | ||
Revision as of 20:43, 21 October 2021
J23100-B0030-LMT-his-GFP-B0010
This part contains a Vibrio natriegens-optimized GFP coding sequence, and the GFP is fused at N-terminal with his-tag in order to let us purify the protein. His-GFP is fused with secretory peptide from LMT.We use BBa_K3739041 to construct the expression system and help prove the function of some signal peptide.
Biology
LMT
Lytic murein transglycosylase (LMT) is an enzyme which is able to degrade murein, a component of cell wall of bacteria. There are two kind of LMTs existing in E.coli: the membrane-binding one and the soluble one. The gene coding LMT homolog is also incorporated in the genome of Vibrio natriegens. The LMT signal peptide (named LMT in our parts) is from the LMT homolog, which can lead the fused protein secreted out of Vibrio natriegens.
GFP
Protein tags are peptides genetically fused to the proteins of interest. There are many different kinds of tags developed for for different purposes. For labeling experiments, normally the proteins of interest are tagged directly by fluorescent tags. Green fluorescent protein (GFP) make it easier to study protein localization, interactions and dynamics when fusing with other peptides and proteins.
Usage
Here, we used BBa_K3739010 to construct the expression system and obtained the composite part BBa_K3739041. We transformed the constructed plasmid on the expression vector pET-28a (+) into V. natriegens to extract the protein. This protein work together with those from BBa_K3739038 and BBa_K3739043 to verify the function of our secretory peptides Aly01 and LMT.
Characterization
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 661
Illegal NgoMIV site found at 1097
Illegal NgoMIV site found at 1832
Illegal NgoMIV site found at 2068
Illegal AgeI site found at 407
Illegal AgeI site found at 497
Illegal AgeI site found at 734
Illegal AgeI site found at 1561
Illegal AgeI site found at 2057 - 1000COMPATIBLE WITH RFC[1000]