Difference between revisions of "Part:BBa K1720000"
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We used lentiviral vector to transfected alpha 3 unit and beta 3 unit to HEK293 cells together with a GFP reporter and in vivo green fluorescence signal was observed under fluorescence microscope, it meant that this part was transfected into HEK293 cells successfully. Then we used qRT-PCR to observe transcriptional level of alpha 3 unit. 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.e detected the cGMP concentration with Elisa Kit.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 Kit to detect cGMP level. The results are as follow: | We used lentiviral vector to transfected alpha 3 unit and beta 3 unit to HEK293 cells together with a GFP reporter and in vivo green fluorescence signal was observed under fluorescence microscope, it meant that this part was transfected into HEK293 cells successfully. Then we used qRT-PCR to observe transcriptional level of alpha 3 unit. 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.e detected the cGMP concentration with Elisa Kit.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 Kit to detect cGMP level. The results are as follow: | ||
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<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
Revision as of 07:00, 26 August 2015
Human guanylate cyclase1,soluble, alpha 3 unit
This is a part used for coding soluble human guanylate cyclase subunit:Alpha 3.Alpha 3 unit interacts with a beta subunit to form the guanylate cyclase enzyme. Soluble guanylate cyclases(sGC) are heterodimeric proteins that catalyze the conversion of GTP to 3',5'-cyclic GMP and pyrophosphate. It is important for smooth muscle relaxation in the cardiovascular system. sGC contains an HNOX domain, which serves as a receptor for ligands such as nitric oxide, oxygen and nitrovasodilator drugs.There are several different isoforms of sGC subunit ,but the cardiovascular system abundant with alpha 3 unit and beta 3 unit.
We used lentiviral vector to transfected alpha 3 unit and beta 3 unit to HEK293 cells together with a GFP reporter and in vivo green fluorescence signal was observed under fluorescence microscope, it meant that this part was transfected into HEK293 cells successfully. Then we used qRT-PCR to observe transcriptional level of alpha 3 unit. 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.e detected the cGMP concentration with Elisa Kit.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 Kit to detect cGMP level. The results are as follow: File:C:\Users\Administrator\Desktop\1-1.jpg Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 1498
Illegal PstI site found at 1827 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 1498
Illegal PstI site found at 1827 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 1498
Illegal PstI site found at 1827 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 1498
Illegal PstI site found at 1827 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1576
Illegal SapI site found at 1446