Difference between revisions of "Part:BBa K3739007"
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===Biology=== | ===Biology=== | ||
Aly01 | Aly01 | ||
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Aly01 is alginate lyase from ''Vibrio natriegens'' SK42.001, which is secreted out and able to digest alginate to unsaturated alginate oligosaccharide. Its signal peptide (named Aly01 in our parts), which is fused with heterogenous protein, is performed well in ''E. coli''. It is implied that the heterogenous protein fused with Aly01 signal peptide may be also secreted efficiently. | Aly01 is alginate lyase from ''Vibrio natriegens'' SK42.001, which is secreted out and able to digest alginate to unsaturated alginate oligosaccharide. Its signal peptide (named Aly01 in our parts), which is fused with heterogenous protein, is performed well in ''E. coli''. It is implied that the heterogenous protein fused with Aly01 signal peptide may be also secreted efficiently. | ||
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GFP | GFP | ||
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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. | 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. | ||
Revision as of 20:32, 21 October 2021
Aly01-his-GFP
Aly01 is a signal peptide from the alginate lyase of Vibrio. natriegens and GFP can show the secretion effciency of Aly01.
Biology
Aly01
Aly01 is alginate lyase from Vibrio natriegens SK42.001, which is secreted out and able to digest alginate to unsaturated alginate oligosaccharide. Its signal peptide (named Aly01 in our parts), which is fused with heterogenous protein, is performed well in E. coli. It is implied that the heterogenous protein fused with Aly01 signal peptide may be also secreted efficiently.
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_K3739003 to construct the expression system and obtained the composite part BBa_K3739043. 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_K3739041 to verify the function of our secretory peptides Aly01 and LMT.
Characterization
1.Colony PCR
References
1.Meng, Q. et al. Characterization and enhanced extracellular overexpression of a new salt-activated alginate lyase. Journal of the science of food and agriculture 101, 5154-5162, doi:10.1002/jsfa.11161 (2021).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 73
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 151