Difference between revisions of "Part:BBa K1720002"
| Line 7: | Line 7: | ||
Before using this part please add a prompter, in our project we add an EF1A promoter to this part.Since EF1A contains Pstl site,we did not add it to this part. | Before using this part please add a prompter, in our project we add an EF1A promoter to this part.Since EF1A contains Pstl site,we did not add it to this part. | ||
| − | This part with a GFP reporter was then transfected into HEK293 cells by lentiviral Vector 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.Finally we detected the cGMP concentration with Elisa Kit. | + | This part with a GFP reporter was then transfected into HEK293 cells by lentiviral Vector 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.Finally we detected the cGMP concentration with Elisa Kit. |
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
Revision as of 05:13, 26 August 2015
Hypoxia-induced promotor
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.
Before using this part please add a prompter, in our project we add an EF1A promoter to this part.Since EF1A contains Pstl site,we did not add it to this part.
This part with a GFP reporter was then transfected into HEK293 cells by lentiviral Vector 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.Finally we detected the cGMP concentration with Elisa Kit.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal SpeI site found at 41
- 12INCOMPATIBLE WITH RFC[12]Illegal SpeI site found at 41
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal SpeI site found at 41
- 25INCOMPATIBLE WITH RFC[25]Illegal SpeI site found at 41
- 1000COMPATIBLE WITH RFC[1000]