Part:BBa_K4960021:Design
Engineered Mitochondrial Uncoupler Pdp1NTD-EGFP-UCP1
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1258
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 235
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
In order to prevent UCP1 from forming inclusion bodies after expression in Escherichia coli and facilitate the localization of UCP1 after transfection, we introduced EGFP protein for fusion expression with UCP1. We designed a flexible peptide linker (GGSGG) to link Pdp1NTD, UCP1 and EGFP to form a fusion protein. However, based on the text result of 砖号, we switched the order of EGFP and UCP1 to ensure that the n-terminal sequence of UCP1 and the Pdp1NTD will no longer interact. (Figure 1) Figure 1. Updated schematic diagram of design ideas. In the process of designing part, we switched the original sequence of EGFP and UCP1, and carried out the same experimental treatment as a new group of experimental groups, hoping to solve the problems encountered before.
Source
E.coli, Photorhabdus