Difference between revisions of "Part:BBa K4140025"
Ahmed Mattar (Talk | contribs) |
(→Part Description) |
||
| (16 intermediate revisions by one other user not shown) | |||
| Line 1: | Line 1: | ||
__NOTOC__ | __NOTOC__ | ||
| − | <partinfo> | + | <partinfo>BBa_K4140025 short</partinfo> |
==Part Description== | ==Part Description== | ||
| − | Coding sequence for an enzyme that causes phenylalanine to be hydroxylated in order to produce tyrosine. PAH is a hydroxylase that needs biopterin. It's linked to KP-SP ISA secreting peptide signal at the N-terminus of the protein to transmit it from intra cellular to | + | It's a Coding sequence for an enzyme that causes phenylalanine to be hydroxylated in order to produce tyrosine. PAH is a hydroxylase that needs biopterin. It's linked to KP-SP ISA secreting peptide signal at the N-terminus of the protein to transmit it from intra-cellular to extracellular |
==Usage== | ==Usage== | ||
| − | We are developing a tunable therapeutic approach to give the body the ability to produce phenylalanine hydroxylase (PAH) by implementing our designed therapeutic circuit in a cell-based system that produces PAH according to phenylalanine and tyrosine levels as shown in figure 1. | + | We are developing a tunable therapeutic approach to give the body the ability to produce phenylalanine hydroxylase (PAH) by implementing our designed therapeutic circuit in a cell-based system that produces PAH according to phenylalanine and tyrosine levels as shown in figure 1. In the presence of phenylalanine (Phe), the TyrR will activate tyrP, accordingly tyrP will enhance the production of PAH gene. While the inhibitory effect of the ParoF will not be initiated, thus L7Ae will be expressed, preventing the expression of the dcas12g. |
| − | + | ||
| + | [[File:imagee.png|Right ]] | ||
| + | |||
| + | Figure 1. (shows the E.coli-based system in our therapeutic platform.) | ||
| + | |||
| + | ==Characterization by mathematical modeling== | ||
| + | We are using mathematical modeling to detect the increased level of phenylalanine (phe) in phenylketonuria patients in our therapeutic platform. It depends on E.coli-based system through a cascade of reactions to finally end by formation of PAH that is deficient in PKU patients as shown in graph (1). | ||
| + | |||
| + | |||
| + | [[File:capture6.png|Right|]] | ||
| + | <br><br><br> | ||
| + | Graph(1) illustrates a direct relation between phenylalanine and PAH ,so as the biomarker increases, the released amount of the enzyme increases till it reaches constant value after about 30 time units. Therefore, the maximum amount of the biomarker releases the maximum amount of PAH. | ||
| + | ==Experimental Characterization== | ||
| + | [[File:tube121.png|right|]] | ||
| + | <br><br><br><br><br><br><br> | ||
| + | This figure shows an experimental characterization of this part as it's validated through gel electrophoresis as it is in lane 10 (the last one). The run part (ordered from IDT) included KP-SP - PAH. | ||
| + | <br><br><br><br><br><br><br><br><br><br><br><br><br><br> | ||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
| Line 14: | Line 29: | ||
<!-- --> | <!-- --> | ||
<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
| − | <partinfo> | + | <partinfo>BBa_K4140025 SequenceAndFeatures</partinfo> |
<!-- Uncomment this to enable Functional Parameter display | <!-- Uncomment this to enable Functional Parameter display | ||
===Functional Parameters=== | ===Functional Parameters=== | ||
| − | <partinfo> | + | <partinfo>BBa_K4140025 parameters</partinfo> |
<!-- --> | <!-- --> | ||
Latest revision as of 04:45, 12 October 2022
PAH releasing device
Part Description
It's a Coding sequence for an enzyme that causes phenylalanine to be hydroxylated in order to produce tyrosine. PAH is a hydroxylase that needs biopterin. It's linked to KP-SP ISA secreting peptide signal at the N-terminus of the protein to transmit it from intra-cellular to extracellular
Usage
We are developing a tunable therapeutic approach to give the body the ability to produce phenylalanine hydroxylase (PAH) by implementing our designed therapeutic circuit in a cell-based system that produces PAH according to phenylalanine and tyrosine levels as shown in figure 1. In the presence of phenylalanine (Phe), the TyrR will activate tyrP, accordingly tyrP will enhance the production of PAH gene. While the inhibitory effect of the ParoF will not be initiated, thus L7Ae will be expressed, preventing the expression of the dcas12g.
Figure 1. (shows the E.coli-based system in our therapeutic platform.)
Characterization by mathematical modeling
We are using mathematical modeling to detect the increased level of phenylalanine (phe) in phenylketonuria patients in our therapeutic platform. It depends on E.coli-based system through a cascade of reactions to finally end by formation of PAH that is deficient in PKU patients as shown in graph (1).
Graph(1) illustrates a direct relation between phenylalanine and PAH ,so as the biomarker increases, the released amount of the enzyme increases till it reaches constant value after about 30 time units. Therefore, the maximum amount of the biomarker releases the maximum amount of PAH.
Experimental Characterization
This figure shows an experimental characterization of this part as it's validated through gel electrophoresis as it is in lane 10 (the last one). The run part (ordered from IDT) included KP-SP - PAH.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 383
Illegal BamHI site found at 910
Illegal XhoI site found at 620 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]