Part:BBa_K3076200

Coding sequence of Vanabin2 gene from Ascidia sydneiensis samea

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.

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.

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.

Assay : Test for copper absorption ability on E.coli transformed with Vanabin

Abstract of experiment

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.

Graph 1: Percentage decrease of copper concentration along time

Graph 1: The percentage decrease of copper concentration in the culture medium

Results

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Future work

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