Difference between revisions of "Part:BBa K3076200"
 
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==Description==
 
==Description==
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<p>This part contains the coding sequence of the <i>vanabin2</i> 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.</p>
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<p><br>This part contains the coding sequence of the <i>vanabin2</i> 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.</br></p>
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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/>
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Although it’s a <i>“vanadium” </i>binding protein, literature reported that copper (II) ions inhibits the binding of <i>vanadium</i> by competing for the binding sites. [1] It showed that VB actually has a higher affinity for copper. Therefore, the <i>VB</i> seems to be an ideal protein for our project to enhance the metal accumulating ability of <i>E. coli</i>.
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==Use and assays==
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==Usage and Biology==
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<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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We synthesized the <i>vanabin2</i> 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==
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===Reference===
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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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[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 Ascidia sydneiensis samea. Applied and Environmental Microbiology, 69(11), 6442–6446. doi: 10.1128/aem.69.11.6442-6446.2003
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==Design notes==
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<span class='h3bb'>Sequence and Features</span>
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Overlapping sequence of pET28a multiple cloning site was added to flank the CDS for cloning by Hifi assembly.
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<partinfo>BBa_K3076200 SequenceAndFeatures</partinfo>
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==Reference==
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[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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Latest revision as of 13:55, 13 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) ions 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.

Usage and Biology

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.

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 Ascidia sydneiensis samea. Applied and Environmental Microbiology, 69(11), 6442–6446. doi: 10.1128/aem.69.11.6442-6446.2003

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 90
  • 1000
    COMPATIBLE WITH RFC[1000]