Difference between revisions of "Part:BBa K1898200"
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==Sequencing==
 
==Sequencing==
  
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We sent GSR out to sequencing after moving them to iGEM BioBrick. 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.  
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We sent GSR out to sequencing after moving it to iGEM BioBrick. 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:
 
Sequence with vf2 primer:

Revision as of 17:46, 18 October 2016


GSR, glutathione reductase

This DNA codes for GSR, also known as glutathione reductase, which catalyzes the conversion of glutathione to glutathione disulfide.

Usage and Biology

GSR is an enzyme that catalyzes the formation of 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. It has been reported that age-induced cataract is mainly caused by the oxidation of the lens protein.[2] The delivery of GSR to the lens should help prevent cataracts from happening.

[1] Sius, H. (n.d.). Glutathione and its role in cellular functions.

[2] Spector, A. (n.d.). Oxidative stress-induced cataract: Mechanism of action.

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 below. 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 after moving it to iGEM BioBrick. 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.