Difference between revisions of "Part:BBa K3076200"
 
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<partinfo>BBa_K3076200 short</partinfo>
 
<partinfo>BBa_K3076200 short</partinfo>
  
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==Description==
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<p><br>This part contains the coding sequence of 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 of the accumulation is still being unsolved.</br></p>
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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>Although it’s a “vanadium” binding protein, literature reported that copper (II) inhibits the binding of vanadium by competing 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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<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 constraint, the work is still in progress.
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<h3>Assay : Test for copper absorption ability on <i>E.coli</i> transformed with Vanabin</h3>
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<h4>Abstract of experiment</h4>
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<p>We utilized pET-151/TOPO vector to drive the expression of Vanabin coding sequence under T7 promoter inside BL21(DE) E. coli strain. Then, the bacteria transformants will be tested inside copper (II) sulphate added culture medium to measure their copper absorption efficiency along time.
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<br><p><u>Graph 1: Percentage decrease of copper concentration along time</u></p>
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<p><u>Graph 1: The percentage decrease of copper concentration in the culture medium
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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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<h3>Results</h3>
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==Usage and Biology==
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<br>.</br>
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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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<h3>Future work</h3>
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===Reference===
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<br>.</br>
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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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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K3076200 SequenceAndFeatures</partinfo>

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]