Difference between revisions of "Part:BBa K1898200"
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==Sequencing== | ==Sequencing== | ||
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Sequence with vr primer: | Sequence with vr primer: | ||
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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. | 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 16:46, 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. Retrieved October 18, 2016, from http://www.sciencedirect.com/science/article/pii/S089158499900177X [2] Spector, A. (n.d.). Oxidative stress-induced cataract: Mechanism of action. Retrieved October 18, 2016, from http://www.ncbi.nlm.nih.gov/pubmed/7672510
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 743
Illegal BamHI site found at 1427 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 28
- 1000INCOMPATIBLE 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).
Sequencing
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.
Sequence with vf2 primer:
Sequence with vr primer:
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.