"
Team:UNIK Copenhagen/Tripartite split GFP
From 2014.igem.org
| Line 25: | Line 25: | ||
table.sequence_description { | table.sequence_description { | ||
| - | |||
| - | |||
| - | |||
background-color: white; | background-color: white; | ||
border-collapse:collapse; | border-collapse:collapse; | ||
| Line 39: | Line 36: | ||
font-family: "Lucida sans unicode", sans-serif; | font-family: "Lucida sans unicode", sans-serif; | ||
font-size: 12px; | font-size: 12px; | ||
| - | margin | + | margin: 3px,3px,3px,3px; |
| - | + | ||
} | } | ||
</style> | </style> | ||
| Line 68: | Line 64: | ||
| - | <table class="sequence_description" id="about_gene" | + | <table class="sequence_description" id="about_gene"></table></div> |
<script> | <script> | ||
| Line 76: | Line 72: | ||
function suc2Function() { | function suc2Function() { | ||
| - | document.getElementById("about_gene").innerHTML=" | + | document.getElementById("about_gene").innerHTML="<p>suc2 codes for a signal sequence that localizes the translation to ER</p>"; |
} | } | ||
function fabFunction() { | function fabFunction() { | ||
| - | document.getElementById("about_gene").innerHTML=" | + | document.getElementById("about_gene").innerHTML="<p>this seguence codes for the FAB fragment</p>"; |
} | } | ||
function gfpFunction() { | function gfpFunction() { | ||
| - | document.getElementById("about_gene").innerHTML=" | + | document.getElementById("about_gene").innerHTML="<p>this gene codes for one of the GFP fragments</p>"; |
} | } | ||
</script> | </script> | ||
Revision as of 11:12, 6 August 2014
TRIPARTITE SPLIT GFP
We split GFP into one large and two small fragments that when put in close-proximity to each other will fuse together into a functional GFP. We then link the two small GFP fragments to antibodies on a genetic level and having the third fragment unbound. Antigens with multiple epitopes can then bind those engineered antibodies, bringing the GFP fragments into close proximity. This will lead to the formation of functional GFP in presence of antigens and the large, unbound GFP part. By cloning this system into yeast (Saccharomyces cerevisiae) and making it secrete all three parts, we will arrive at a new single step detection tool where a sample, mixed with water and yeast, will prove the presence of a targeted pathogen by an increase in fluorescence. Once a stable yeast strain is obtained, targeting a specific antigen, simple scale up would allow for mass production and chemical usage minimized. Therefore, this tool should be both cheap and environmentally responsible compared to existing detection methods such as ELISA. The simplicity of this system would allow for dried yeast to be send across the world for easy field-testing.
Our split-GFP approach was inspired by the work of (Cabantous et al., 2013) who used it for protein-protein interactions. Realising that if we replaced the interacting proteins with antibodies with close binding sites, an antigen detector would be possible.
References:
Cabantous, S., Nguyen, H. B., Pedelacq, J.-D., Koraïchi, F., Chaudhary, A., Ganguly, K., … Waldo, G. S. (2013). A new protein-protein interaction sensor based on tripartite split-GFP association. Scientific Reports, 3, 2854. doi:10.1038/srep02854
We are using homolog recombination to transform the yeast strains.
Gene 1:
Touch the lego bricks to see what sequences the gene consist of and click on the sequences to read more about their function.
