Difference between revisions of "Team:Utrecht/Improve"
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<div class = "customelementM5A" id = "Introduction"> | <div class = "customelementM5A" id = "Introduction"> | ||
| − | <p> In order to measure the activity of custom receptors we set up an assay that measures the activity of hybrid receptors between the copper sensing receptor protein CusS and Tar by means of RFP expression. When bacteria are cultured in presence of aspartate, RFP expression is induced by the phosphorylation and subsequent binding of CusR to the copper responsive promoter element CopA. We opted to use BBa_K1980004<a class = "url_icon" href = "http://parts.igem.org/Part:BBa_K1980004"></a> which contains the sequence of the CusS/CusR responsive promoter that lacks a Ribosome binding site. We designed a version of this part that is codon optimized for <i>E. coli | + | <p> In order to measure the activity of custom receptors we set up an assay that measures the activity of hybrid receptors between the copper sensing receptor protein CusS and Tar by means of RFP expression. When bacteria are cultured in presence of aspartate, RFP expression is induced by the phosphorylation and subsequent binding of CusR to the copper responsive promoter element CopA. We opted to use BBa_K1980004<a class = "url_icon" href = "http://parts.igem.org/Part:BBa_K1980004"></a> which contains the sequence of the CusS/CusR responsive promoter that lacks a Ribosome binding site. We designed a version of this part that is codon optimized for <i>E. coli</i> DH5α and contains a ribosome binding site. We tested the functionality of this part in several ways and show that it is a functional part that responds to part BBa_K2736109<a class = "url_icon" href = "http://parts.igem.org/Part:BBa_K2736109"></a>.</p> |
</div> | </div> | ||
Latest revision as of 19:44, 17 October 2018
In order to measure the activity of custom receptors we set up an assay that measures the activity of hybrid receptors between the copper sensing receptor protein CusS and Tar by means of RFP expression. When bacteria are cultured in presence of aspartate, RFP expression is induced by the phosphorylation and subsequent binding of CusR to the copper responsive promoter element CopA. We opted to use BBa_K1980004 which contains the sequence of the CusS/CusR responsive promoter that lacks a Ribosome binding site. We designed a version of this part that is codon optimized for E. coli DH5α and contains a ribosome binding site. We tested the functionality of this part in several ways and show that it is a functional part that responds to part BBa_K2736109.