Difference between revisions of "Part:BBa K1720001:Design"

 
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===Design Notes===
 
===Design Notes===
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We used lentiviral vector to transfected alpha 3 unit and beta 3 unit to HEK293 cells together. Since soluble guanylate cyclases (sGC) are heterodimeric proteins that catalyze the conversion of GTP to 3',5'-cyclic GMP(cGMP) and pyrophosphate. The level of cGMP will be up regulated.The positive control was HEK293 cells that treat with Sodium Nitroprusside ,a NO donator that activate sGC and up regulate the level of cGMP. A negative control was made by transfecting an empty vector that does not contain sGC subunit. We used Elisa to detect cGMP level.
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The vector we used in our experiment was created using scarless golden gate assembly.We cloned this basic part into a psb1c3 vector without promoter , so that user can choose their own promoter for different purpose. After standardizing our part, a promoter can be easily inserted in front of our part. This part is only a subunit of human guanylate cyclase,.To form holoenzyme this part needs to work with BBa_K1720000 together.
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===Source===
 
===Source===
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===References===
 
===References===
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1. Gheorghiade M, Marti CN, Sabbah HN, Roessig L, Greene SJ, Bohm M, Burnett JC, Campia U, Cleland JG, Collins SP et al: Soluble  guanylate cyclase: a potential therapeutic target for heart failure. Heart failure reviews 2013, 18(2):123-134.
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2. Sharina IG, Cote GJ, Martin E, Doursout MF, Murad F: RNA splicing in regulation of nitric oxide receptor soluble guanylyl cyclase. Nitric oxide : biology and chemistry / official journal of the Nitric Oxide Society 2011, 25(3):265-274.
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3. Marinko M, Novakovic A, Nenezic D, Stojanovic I, Milojevic P, Jovic M, Ugresic N, Kanjuh V, Yang Q, He GW: Nicorandil directly and cyclic GMP-dependently opens K channels in human bypass grafts. Journal of pharmacological sciences 2015.

Latest revision as of 07:32, 4 September 2015


Human guanylate cyclase1,soluble, beta 3 unit


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 579
    Illegal BamHI site found at 800
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The vector we used in our experiment was created using scarless golden gate assembly.We cloned this basic part into a psb1c3 vector without promoter , so that user can choose their own promoter for different purpose. After standardizing our part, a promoter can be easily inserted in front of our part. This part is only a subunit of human guanylate cyclase,.To form holoenzyme this part needs to work with BBa_K1720000 together.

Source

We get the informations from this link: http://www.ncbi.nlm.nih.gov/gene/2983

References

1. Gheorghiade M, Marti CN, Sabbah HN, Roessig L, Greene SJ, Bohm M, Burnett JC, Campia U, Cleland JG, Collins SP et al: Soluble guanylate cyclase: a potential therapeutic target for heart failure. Heart failure reviews 2013, 18(2):123-134.

2. Sharina IG, Cote GJ, Martin E, Doursout MF, Murad F: RNA splicing in regulation of nitric oxide receptor soluble guanylyl cyclase. Nitric oxide : biology and chemistry / official journal of the Nitric Oxide Society 2011, 25(3):265-274.

3. Marinko M, Novakovic A, Nenezic D, Stojanovic I, Milojevic P, Jovic M, Ugresic N, Kanjuh V, Yang Q, He GW: Nicorandil directly and cyclic GMP-dependently opens K channels in human bypass grafts. Journal of pharmacological sciences 2015.