Difference between revisions of "Part:BBa K2520008"
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This device is a complete system for displaying proteins on cells' membrane. We combined three independent systems into one, which allows for the expression of three unique proteins. The Igk leader (BBa_K2520024,BBa_K2520028,BBa_K2520029) is a short signal peptide that prompts the translocation of a protein to the cellular membrane. PDGFR (BBa_K2520026,BBa_K2520036,BBa_K2520037) is a trans-membrane domain that anchors all the components located between the igk leader and the PDGFR itself to the membrane. HA (BBa_K2520030), Myc (BBa_K2520031) and His (BBa_K2520032) are tags that bind to specific antibodies and provide an indirect method for verifying the display of the proteins on the membrane. The proteins that we chose to express on the membrane are three epitopes that are known targets of multiple sclerosis (BBa_K2520038,BBa_K2520039,BBa_K2520040) and are components related to our specific project. P2A (BBa_K2520033) and T2A (BBa_K2520034) are internal peptide cleavage sites that allow for the equimolar expression of multiple proteins under the same promoter. | This device is a complete system for displaying proteins on cells' membrane. We combined three independent systems into one, which allows for the expression of three unique proteins. The Igk leader (BBa_K2520024,BBa_K2520028,BBa_K2520029) is a short signal peptide that prompts the translocation of a protein to the cellular membrane. PDGFR (BBa_K2520026,BBa_K2520036,BBa_K2520037) is a trans-membrane domain that anchors all the components located between the igk leader and the PDGFR itself to the membrane. HA (BBa_K2520030), Myc (BBa_K2520031) and His (BBa_K2520032) are tags that bind to specific antibodies and provide an indirect method for verifying the display of the proteins on the membrane. The proteins that we chose to express on the membrane are three epitopes that are known targets of multiple sclerosis (BBa_K2520038,BBa_K2520039,BBa_K2520040) and are components related to our specific project. P2A (BBa_K2520033) and T2A (BBa_K2520034) are internal peptide cleavage sites that allow for the equimolar expression of multiple proteins under the same promoter. | ||
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| − | The CMV promoter (BBa_K1119006) is a constitutive expression promoter in mammalian cells and hGH (BBa_K404108) serves as a terminator. | + | The CMV promoter (BBa_K1119006) is a constitutive expression promoter in mammalian cells and hGH (BBa_K404108) serves as a terminator. The structure of this device is shown below. |
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===Structure=== | ===Structure=== | ||
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===Usage and Biology=== | ===Usage and Biology=== | ||
| + | We characterized this device by transfecting HEK-293 cells with this device and two more variations of it, with pDisplay backbone including epitope of bee venom PLA antigen as a positive control, with a plasmid containing CMV promoter and GFP as a reporter gene as a negative control and also non-transfected cells. We incubate the transfected cells with specific antibodies that bind to three different tags (HA, Myc and His) conjugated to chromophores and measured the fluorescence intensity emitted. The device described above is the variation that has the highest fluorescence intensity of all three variations, comparing to the positive control (Figure 1). | ||
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Revision as of 12:23, 26 October 2017
Status: 500
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For help, please send mail to this site's webmaster, giving this error message and the time and date of the error.
This device is a complete system for displaying proteins on cells' membrane. We combined three independent systems into one, which allows for the expression of three unique proteins. The Igk leader (BBa_K2520024,BBa_K2520028,BBa_K2520029) is a short signal peptide that prompts the translocation of a protein to the cellular membrane. PDGFR (BBa_K2520026,BBa_K2520036,BBa_K2520037) is a trans-membrane domain that anchors all the components located between the igk leader and the PDGFR itself to the membrane. HA (BBa_K2520030), Myc (BBa_K2520031) and His (BBa_K2520032) are tags that bind to specific antibodies and provide an indirect method for verifying the display of the proteins on the membrane. The proteins that we chose to express on the membrane are three epitopes that are known targets of multiple sclerosis (BBa_K2520038,BBa_K2520039,BBa_K2520040) and are components related to our specific project. P2A (BBa_K2520033) and T2A (BBa_K2520034) are internal peptide cleavage sites that allow for the equimolar expression of multiple proteins under the same promoter.
The CMV promoter (BBa_K1119006) is a constitutive expression promoter in mammalian cells and hGH (BBa_K404108) serves as a terminator. The structure of this device is shown below.
Structure
Sequence and Features Status: 500 Content-type: text/html
Software error:
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