Difference between revisions of "Part:BBa K1898200"
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==Gel Pictures==
 
==Gel Pictures==
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We designed primers synthesized by Tri-I Biotech to remove stop codon and to move GSR cDNA to iGEM BioBrick. PCR was set up and the gel picture is shown above. The expected size for GSR PCR check is ~1.8kb, which is shown in lane__:
+
We designed primers to remove stop codon and to move GSR cDNA to iGEM BioBrick. PCR was set up and the gel picture is shown above. Lane 1 is 1kb ladder. The expected size for GSR PCR check is ~1.8kb, which is shown in lane 5-8 (boxed in red).
 +
 
 +
https://static.igem.org/mediawiki/parts/b/bd/Gsr_gel.png
  
  
 
==Sequencing==
 
==Sequencing==
  
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The sequencing result shows that the GSR sequencing result is correct (with no stop codon):
+
We sent GSR out to sequencing. The sequencing results are unable to confirm the sequence from 480 to 744 bp. The four cutting sites are highlighted in yellow and GSR is highlighted in orange.
 +
 
 +
Sequence with vf2 primer:
 +
https://static.igem.org/mediawiki/parts/f/ff/Gsr_vf2.png
 +
 
 +
Sequence with vr primer:
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https://static.igem.org/mediawiki/parts/4/4f/Gsr_vr.png
 +
 
 +
This GSR is later used in BBa_K1898250, which was also sent to sequencing. From the sequencing file snipped in part.igem.org/Part:BBa_K1898250, the GSR sequence from 480 to 744 bps is confirmed to be correct.

Revision as of 09:28, 18 October 2016


GSR, glutathione reductase

GSR codes for gluatathione reductase, an enzyme that catalyzes the conversion of glutathione reductase to glutathione. Glutathione is an antioxidant exists in humans, plants, fungi, animal that prevents cellular damage by reactive oxygen species. [1] In our project, we aim to deliver GSR to the lens, where GSH will be produced and prevent reactive oxygen species from damaging the lens proteins. Age-induced cataract is mainly caused by reactive oxygen species oxidation of the lens protein. The increase in GSH will help prevent cataracts from happening.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 743
    Illegal BamHI site found at 1427
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 28
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 1138
    Illegal SapI.rc site found at 1546


Gel Pictures

We designed primers to remove stop codon and to move GSR cDNA to iGEM BioBrick. PCR was set up and the gel picture is shown above. Lane 1 is 1kb ladder. The expected size for GSR PCR check is ~1.8kb, which is shown in lane 5-8 (boxed in red).

Gsr_gel.png


Sequencing

We sent GSR out to sequencing. The sequencing results are unable to confirm the sequence from 480 to 744 bp. The four cutting sites are highlighted in yellow and GSR is highlighted in orange.

Sequence with vf2 primer: Gsr_vf2.png

Sequence with vr primer: Gsr_vr.png

This GSR is later used in BBa_K1898250, which was also sent to sequencing. From the sequencing file snipped in part.igem.org/Part:BBa_K1898250, the GSR sequence from 480 to 744 bps is confirmed to be correct.