Team:Hong Kong JSS/Contribution
Characterisation of previous part
Since our project look for a method to increase the metal absorption ability of E. coli, we have previously identified the metal-binding protein - metallothionein (MT) from several different organisms.
One of the candidate in our list that showed high affinity in binding different metal ions was the metallothionein from Mycobacterium tuberculosis (MyMT). And we found that the gene code for MyMT had previously been cloned and tested by the 2016 Oxford team (BBa_K1980002) and the protein was used by them to chelate copper for treating Wilson's Disease. Their result showed MyMT could indeed bind copper ions and so that we designed to see if MyMT can also increase the copper absorption and binding ability in E. coli.
We synthesized the part with a strong constitutive promoter (BBa_J23100), RBS (BBa_B0032) and a double terminator (BBa_B0015) by IDT gBlock and then assembled it into pSB1C3 vector. The plasmid was then transformed into E. coli for expression and copper absorption assay. The result of the assay had been documented to the MyMT parts registry Main Page. For details, please refer to our Resultpage.
We synthesized the part with a strong constitutive promoter (BBa_J23100), RBS (BBa_B0032) and a double terminator (BBa_B0015) by IDT gBlock and then assembled it into pSB1C3 vector. The plasmid was then transformed into E. coli for expression and copper absorption assay. The result of the assay had been documented to the MyMT parts registry Main Page. For details, please refer to our Resultpage.
In our experiment, Mymt transformant can reduced the copper level in water by 11.4% after 4 hours. The reduction is significantly different from the control.