Difference between revisions of "Part:BBa K3610018"

Latest revision as of 23:03, 27 October 2020

mCherry N-terminal - codon optimized for C. reinhardtii

This part is a modified version of Part:BBa_K1093016. It has been codon optimized for expression in C. reinhardtii and additionally the split was made at amino acids 158 and 159.

Function of the part

mCherry is a monomeric red fluorescent protein that has been used for observing components by fusing the sequence to a protein of interes and observing the mCherry by flourescent spectroscopy and microscopy. The mCherry protein is the basis for the bimolecular fluorescence fusion technique which is used to visualize protein-protein interaction. The mCherry is split into two parts, in our case between amino acids 158 and 159. The two components can be fused to proteins of interest and, upon dimerization, are reconstituted to the fully functional red fluorescent protein (Excitation/Emission wavelengths: 587/610 nm).

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 5: / Line 5: @@
 This part is a modified version of [[Part:BBa_K1093016]]. It has been codon optimized for expression in C. reinhardtii and additionally the split was made at amino acids 158 and 159.
-===Biology and Usage===
+===Function of the part===
 mCherry is a monomeric red fluorescent protein that has been used for observing components by fusing the sequence to a protein of interes and observing the mCherry by flourescent spectroscopy and microscopy.
 The mCherry protein is the basis for the bimolecular fluorescence fusion technique which is used to visualize protein-protein interaction.
 The mCherry is split into two parts, in our case between amino acids 158 and 159. The two components can be fused to proteins of interest and, upon dimerization, are reconstituted to the fully functional red fluorescent protein (Excitation/Emission wavelengths: 587/610 nm).
-This part contains the N-terminal part of the split mCherry system and can be used together with [[Part:BBa_K3610019]] for investigation of protein-protein interaction in C. reinhardtii.
-In our project, we used this part for visualizing the interaction of leucine-rich repeat receptor-like kinases (LRR-RLKs). We fused the protein to the ectodomains of plant pattern-recognition recpetors which need dimerization to initiate an immune response in plants.
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