<h3id="2.2"class="anchor">2.2) Protein production</h3>
<h5>Production of CBD- and SB7-tagged Proteins and Non-Tagged sfGFP</h5>
<p>After we successfully produced our level 1 JUMP assemblies for our CBD- and SB7-tagged proteins (SB7-sfGFP, CBD-Metallothionein (MT) and CBD-sfGFP) We also produced non-tagged sfGFP to use as controls for the fluorescence assays for assessing the performance of the CBD and SB7 tags. We then transformed them into E. coli BL21(DE3) for protein expression. After protein expression in the BL21(DE3) cell cultures, the cultures were lysed by sonication, and the lysates were run on an SDS-PAGE gel to confirm the presence of our CBD- and SB7-tagged proteins (Figure 10). This SDS-PAGE step also serves as a solubility test to confirm that all of our desired proteins are in the soluble fraction and can be properly incorporated into protein immobilisation methods.</p>
<p>After we successfully produced our level 1 JUMP assemblies for our CBD- and SB7-tagged proteins (SB7-sfGFP, CBD-Metallothionein (MT) and CBD-sfGFP) We also produced non-tagged sfGFP to use as controls for the fluorescence assays for assessing the performance of the CBD and SB7 tags. We then transformed them into <i>E. coli</i> BL21(DE3) for protein expression. After protein expression in the BL21(DE3) cell cultures, the cultures were lysed by sonication, and the lysates were run on an SDS-PAGE gel to confirm the presence of our CBD- and SB7-tagged proteins (Figure 10). This SDS-PAGE step also serves as a solubility test to confirm that all of our desired proteins are in the soluble fraction and can be properly incorporated into protein immobilisation methods.</p>
