Difference between revisions of "Part:BBa K500001"
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Group: iGEM19_Uppsala_Universitet | Group: iGEM19_Uppsala_Universitet | ||
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+ | Author: Jonas Gockel | ||
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+ | Summary: We studied the expression of <partinfo>BBa_K500001</partinfo> in <I> Pichia pastoris </I> under the control of the AOX1 promoter <partinfo>BBa_K3105675</partinfo> and an N-terminal fused α-secretion factor <partinfo>BBa_K3105674</partinfo>. | ||
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+ | To express <partinfo>BBa_K500001</partinfo>, overhang PCR was performed and the gene was cloned into the commercial pPICZαB vector by gibson assembly. In this vector, the gene is controlled by the AOX1-promoter and is cloned in-frame to an N-terminal α-secretion factor from <I>S. cerevisiae</I>. MnP was expressed in the X-33 wildtype strain of <I>Pichia pastoris</I>, where secretion was observed (Figure 1). | ||
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Revision as of 13:48, 18 October 2019
lignin degradation 2
Yeast codon optimization , no terminator codon, from Phanerochaete chrysosporium. Synthetized by Geneart Mn peroxidases (MnP) are extracellular hemeproteins first discovered in the white-rot fungus Phanerochaete chrysosporium, and have been virtually detected in all lignin degrading fungi so far studied. The catalytic cycle of MnP is similar to that of other plant and fungal peroxidases
Contribution
Group: iGEM19_Uppsala_Universitet
Author: Jonas Gockel
Summary: We studied the expression of BBa_K500001 in Pichia pastoris under the control of the AOX1 promoter BBa_K3105675 and an N-terminal fused α-secretion factor BBa_K3105674.
To express BBa_K500001, overhang PCR was performed and the gene was cloned into the commercial pPICZαB vector by gibson assembly. In this vector, the gene is controlled by the AOX1-promoter and is cloned in-frame to an N-terminal α-secretion factor from S. cerevisiae. MnP was expressed in the X-33 wildtype strain of Pichia pastoris, where secretion was observed (Figure 1).
The full expression circuit can be found here:
BBa_K3105682
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 220
Illegal BglII site found at 512 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]