Difference between revisions of "Part:BBa K3076200"
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<p><br>Although it’s a “vanadium” binding protein, literature reported that copper (II) inhibits the binding of vanadium by competing for the binding sites. [1] It showed that VB actually has a higher affinity for copper. Therefore, the VB seems to be an ideal protein for our project to enhance the metal accumulating ability of E. coli.</br><p/> | <p><br>Although it’s a “vanadium” binding protein, literature reported that copper (II) inhibits the binding of vanadium by competing for the binding sites. [1] It showed that VB actually has a higher affinity for copper. Therefore, the VB seems to be an ideal protein for our project to enhance the metal accumulating ability of E. coli.</br><p/> | ||
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==Use and assays== | ==Use and assays== | ||
<p><br>We synthesized the vanabin2 CDS fragment by IDT gBlock and planned to assemble the fragment into pET28a expression vector by Hifi assembly. However, due to time constraints, the work is still in progress. | <p><br>We synthesized the vanabin2 CDS fragment by IDT gBlock and planned to assemble the fragment into pET28a expression vector by Hifi assembly. However, due to time constraints, the work is still in progress. | ||
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==Source of the part== | ==Source of the part== | ||
This part contains the coding sequence of the <i>vanabin2</i> gene. It was identified from the literature review [1] and extracted from NCBI (ACCESSION: AB088204) starting from start codon to stop codon. Then the sequence was codon-optimized for <i>E. coli</i> by the ThermoFisher GeneArt platform. | This part contains the coding sequence of the <i>vanabin2</i> gene. It was identified from the literature review [1] and extracted from NCBI (ACCESSION: AB088204) starting from start codon to stop codon. Then the sequence was codon-optimized for <i>E. coli</i> by the ThermoFisher GeneArt platform. | ||
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==Design notes== | ==Design notes== | ||
Overlapping sequence of pET28a multiple cloning site was added to flank the CDS for cloning by Hifi assembly. | Overlapping sequence of pET28a multiple cloning site was added to flank the CDS for cloning by Hifi assembly. | ||
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==Reference== | ==Reference== | ||
[1] Ueki, T., Sakamoto, Y., Yamaguchi, N., & Michibata, H. (2003). Bioaccumulation of Copper Ions by Escherichia coli Expressing Vanabin Genes from the Vanadium-Rich Ascidian <i>Ascidia sydneiensis samea</i>. Applied and Environmental Microbiology, 69(11), 6442–6446. doi: 10.1128/aem.69.11.6442-6446.2003 | [1] Ueki, T., Sakamoto, Y., Yamaguchi, N., & Michibata, H. (2003). Bioaccumulation of Copper Ions by Escherichia coli Expressing Vanabin Genes from the Vanadium-Rich Ascidian <i>Ascidia sydneiensis samea</i>. Applied and Environmental Microbiology, 69(11), 6442–6446. doi: 10.1128/aem.69.11.6442-6446.2003 | ||
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Revision as of 12:40, 11 October 2019
Coding sequence of Vanabin2 gene from Ascidia sydneiensis samea
Description
This part contains the coding sequence of the vanabin2 gene which encodes a vanadium-binding protein (VB). The gene was identified from a vanadium-rich sea squirt and it has a function of helping the organism to accumulate metal ion for physiological purpose. The reason for the accumulation is still unsolved.
Although it’s a “vanadium” binding protein, literature reported that copper (II) inhibits the binding of vanadium by competing for the binding sites. [1] It showed that VB actually has a higher affinity for copper. Therefore, the VB seems to be an ideal protein for our project to enhance the metal accumulating ability of E. coli.
We synthesized the vanabin2 CDS fragment by IDT gBlock and planned to assemble the fragment into pET28a expression vector by Hifi assembly. However, due to time constraints, the work is still in progress.