Difference between revisions of "Part:BBa K3196032"
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We combine pelA, SLAC genes, which each adds His tag behind together.Through the previous experiment, we find the signal peptide which is most suitable for the target genes, and form the final pathway.Besides we add an alkali peptide to maintain pH. | We combine pelA, SLAC genes, which each adds His tag behind together.Through the previous experiment, we find the signal peptide which is most suitable for the target genes, and form the final pathway.Besides we add an alkali peptide to maintain pH. | ||
| + | [[File:T--HUST-China--2019-the -whole-pathway2.jpg|400px|thumb|center| Figure1 path way]] | ||
<h1>'''Characterization'''</h1> | <h1>'''Characterization'''</h1> | ||
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We put pectinase pelA at the first place because the banana fiber is covered by pectin. After dealing with the surface of the fiber, SLAC is as follow to degrade lignin inside the banana further, aiming to gain higher quality of the fiber. Since all these three enzymes’ optimum pH is near 5, we designed a pH system. We use Pgas, a promoter activate at pH equals 5, matching the optimum pH of the enzymes. In case the whole system’s pH decreases too much, we use Pgpda, a promoter activate at pH equals 2, connecting it with an alkali peptide to stabilize pH. | We put pectinase pelA at the first place because the banana fiber is covered by pectin. After dealing with the surface of the fiber, SLAC is as follow to degrade lignin inside the banana further, aiming to gain higher quality of the fiber. Since all these three enzymes’ optimum pH is near 5, we designed a pH system. We use Pgas, a promoter activate at pH equals 5, matching the optimum pH of the enzymes. In case the whole system’s pH decreases too much, we use Pgpda, a promoter activate at pH equals 2, connecting it with an alkali peptide to stabilize pH. | ||
| − | [[File:T--HUST--China--2019-path way 2.png|400px|thumb|center| Figure | + | [[File:T--HUST--China--2019-path way 2.png|400px|thumb|center| Figure 2 ]] |
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
Latest revision as of 13:39, 20 October 2019
pGAP-Kozak-PHO1-pelA-His tag-TT-Pgpda-Kozak-FLO10-SLAC-His tag-TT-Pgas-Alkali-TT
We combine pelA, SLAC genes, which each adds His tag behind together.Through the previous experiment, we find the signal peptide which is most suitable for the target genes, and form the final pathway.Besides we add an alkali peptide to maintain pH.
Characterization
This is a three section part for degrading banana fiber part.
Usage and Biology
We put pectinase pelA at the first place because the banana fiber is covered by pectin. After dealing with the surface of the fiber, SLAC is as follow to degrade lignin inside the banana further, aiming to gain higher quality of the fiber. Since all these three enzymes’ optimum pH is near 5, we designed a pH system. We use Pgas, a promoter activate at pH equals 5, matching the optimum pH of the enzymes. In case the whole system’s pH decreases too much, we use Pgpda, a promoter activate at pH equals 2, connecting it with an alkali peptide to stabilize pH.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1308
Illegal BamHI site found at 478
Illegal BamHI site found at 2127
Illegal BamHI site found at 2138
Illegal BamHI site found at 2149
Illegal BamHI site found at 3501 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 2588
Illegal SapI site found at 238