Difference between revisions of "Part:BBa K4390027"

Revision as of 14:33, 12 October 2022

SB7-tagged sfGFP

This part is not compatible with BioBrick RFC10 assembly but is compatible with the iGEM Type IIS Part standard which is also accepted by iGEM.

Used as a control in our silica immobilisation experiment.

Usage and Biology

SB7 is a 7-amino-acid silica-binding motif from the spore coat protein CotB1 of Bacillus cereus. According to studies, its three arginine residues are important for binding to silica surfaces. We tagged this sequence to the N-terminus of sfGFP. Therefore, when the lysate from a cell culture expressing SB7-tagged sfGFP is mixed with silica beads followed by washing off the supernatant, we would be left with green-coloured silica beads which indicates that sfGFP is bound to them. This is one of the three types of silica-binding tags we explored for immobilising proteins to silica beads.

The green-fluorescent recombinant protein was used as an indicator to show qualitatively and quantitatively whether the SB7 peptide has affinity towards silica beads. This was assayed by comparing the green fluorescence intensity between the initial lysate and the supernatant after addition of the silica beads. There should be a significant reduction in fluorescence intensity to prove that the SB7 peptide indeed binds to silica beads and thus, immobilises the sfGFP fused to it.

Sequence and Features