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To use GFP for <i>C.elegans</i>, we needed to modify and confirm this GFP. After getting sequence of <html><a href="http://parts.igem.org/Part:BBa_K309016">BBa_K309016</a></html> sequence, we find that the part doesn’t start with start codon “ATG". We optimize it by deleting 9bp in front of the start codon. Then the coding sequence starts with “ATG”(Figure 2). | To use GFP for <i>C.elegans</i>, we needed to modify and confirm this GFP. After getting sequence of <html><a href="http://parts.igem.org/Part:BBa_K309016">BBa_K309016</a></html> sequence, we find that the part doesn’t start with start codon “ATG". We optimize it by deleting 9bp in front of the start codon. Then the coding sequence starts with “ATG”(Figure 2). | ||
− | {{SUSTech_Image_Center_8| filename=T--SUSTech_Shenzhen--Charicterization2.png|width=1000px|caption=<B>Figure 2. Alignment between BBa_K309016 and <html><a href="http://parts.igem.org/Part:BBa_K2492000">BBa_K2492000</a></html> (optimized GFP) at start codon.</B>}} | + | {{SUSTech_Image_Center_8| filename=T--SUSTech_Shenzhen--Charicterization2.png|width=1000px|caption=<B>Figure 2. Alignment between <html><a href="http://parts.igem.org/Part:BBa_K309016">BBa_K309016</a></html> and <html><a href="http://parts.igem.org/Part:BBa_K2492000">BBa_K2492000</a></html> (optimized GFP) at start codon.</B>}} |
− | We successfully improved characterization of GFP for <i>C.elegans</i>. We submit this optimized GFP as our part <html>< | + | We successfully improved characterization of GFP for <i>C.elegans</i>. We submit this optimized GFP as our part <html><a href="http://parts.igem.org/Part:BBa_K2492000">BBa_K2492000</a></html>. We constructed a Str-1::Chrimson::GEM-GECO::GFP fusion gene and injected it into <i>C.elegans</i>. Promoter Str-1 drives the expression of GFP in neuron AWB, which indicate the AWB location. |
We capture the image of GFP specifically expressed in AWB with confocal microscope and clearly observe AWB with its dendrite(Figure.3). | We capture the image of GFP specifically expressed in AWB with confocal microscope and clearly observe AWB with its dendrite(Figure.3). |
Revision as of 03:31, 2 November 2017
Characterization
Parts
We tried to utilize GFP from biobricks distribution to genetically label neuron AWB in C. elegans’ (Caenorhabditis elegans) head. The biobricks part that specifically marked for C. elegans is the BBa_K309016 submitted by iGEM10_Queens-Canada . Unfortunately, the sample status shows that ”it’s complicated”, which means the sample may not have been sequenced or there is some problems on it (Figure 1).
To use GFP for C.elegans, we needed to modify and confirm this GFP. After getting sequence of BBa_K309016 sequence, we find that the part doesn’t start with start codon “ATG". We optimize it by deleting 9bp in front of the start codon. Then the coding sequence starts with “ATG”(Figure 2).
We successfully improved characterization of GFP for C.elegans. We submit this optimized GFP as our part BBa_K2492000. We constructed a Str-1::Chrimson::GEM-GECO::GFP fusion gene and injected it into C.elegans. Promoter Str-1 drives the expression of GFP in neuron AWB, which indicate the AWB location.
We capture the image of GFP specifically expressed in AWB with confocal microscope and clearly observe AWB with its dendrite(Figure.3).