Part:BBa_K656010:Experience

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Applications of BBa_K656010

The Brown-Stanford iGEM Team 2011 utilized this part in conjunction with the cscB sucrose permease and gfpmut3b parts (see part BBa_K656012 for full construct). We used the gfp reporter to characterized the functionality of this promoter part independent of the functionality of our cscB part (BBa_K656011). The micrographs below demonstrate successful conjugal transfer of our plasmid into Anabaena 7120 and proper functionality of our promoter:

The following series of micrographs is from our initial discovery of successful conjugation, prior to selection:

This stand-alone image shows a very predominant GFP presence in a successful transformant, with faint fluorescence characteristic of negative controls apparent in the background containing other cells in the culture that did not receive the plasmid from the donor e.coli strains. The arrows point to examples of what we believe to be heterocysts within the organism that appear closer to the background level of fluorescence. This is expected because the pSac promoter is specific for vegetative cells.

This series of three images shows the same sample section under different filters: the first is a bright-field image, the second shows chlorophyll fluorescence, and the third is filtered for GFP. Interestingly, the introduction of our plasmid and/or the expression of GFP seems to have altered the amount of chlorophyll production in the organism.

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