Difference between revisions of "Part:BBa K2810009"

Latest revision as of 13:48, 28 September 2018

mCherry, codon optimised for Nicotiana benthamiana and GoldenGate compatible

The sequence of mCherry, derived from part BBa_K133055, but codon optimised for Nicotiana benthamiana . It has extra 5' and 3' sites that make it GoldenGate Compatible. This is because there were no RFPs or derivatives in the iGEM parts registry that were GoldenGate compatible. This part expands the phytobrick registry and can be a powerful tool for plant cell imaging. RFP and its derivatives fluoresce in a different wavelength of light relative to GFP. While chlorophyll autofluoresces in the red region of light (about 680nm), it still has much stronger expression. It is also possible that the photon imager used to measure output detects green light that is simply not absorbed by the chlorophyll. mCherry functions as a monomer, rather than a dimer like GFP, and consequently often has more consistent and higher levels of expression.

This is an improved part. The coding sequence has 5' AATG 3' GCTT 4nt overhangs that makes the sequence conforms to the [http://2016.igem.org/Resources/Plant_Synthetic_Biology/PhytoBricks Phytobrick] golden gate standard. The part can be cloned into a higher order level 1 composite clone using BsaI.

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]

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 <partinfo>BBa_K2810009 short</partinfo>
-The sequence of mCherry, derived from part [https://parts.igem.org/Part:BBa_K133055 BBa_K133055], but codon optimised for <i> Nicotiana benthamiana </i>. It has extra 5' and 3' sites that make it GoldenGate Compatible. This is because there were no RFPs or derivatives in the iGEM parts registry that were GoldenGate compatible. This part expands the phytobrick registry and can be a powerful tool for plant cell imaging. RFP and its derivatives fluoresce in a different wavelength of light relative to GFP, which fluoresces in a similar wavelength to chlorophyll, and thereby is unfavourable to use a reporter gene in plants. mCherry also functions as a monomer, rather than a dimer like GFP, and consequently often has more consistent and higher levels of expression.
+The sequence of mCherry, derived from part [https://parts.igem.org/Part:BBa_K133055 BBa_K133055], but codon optimised for <i> Nicotiana benthamiana </i>. It has extra 5' and 3' sites that make it GoldenGate Compatible. This is because there were no RFPs or derivatives in the iGEM parts registry that were GoldenGate compatible. This part expands the phytobrick registry and can be a powerful tool for plant cell imaging. RFP and its derivatives fluoresce in a different wavelength of light relative to GFP. While chlorophyll autofluoresces in the red region of light (about 680nm), it still has much stronger expression. It is also possible that the photon imager used to measure output detects green light that is simply not absorbed by the chlorophyll. mCherry functions as a monomer, rather than a dimer like GFP, and consequently often has more consistent and higher levels of expression.
 This is an improved part. The coding sequence has 5' AATG 3' GCTT 4nt overhangs that makes the sequence conforms to the [http://2016.igem.org/Resources/Plant_Synthetic_Biology/PhytoBricks Phytobrick] golden gate standard. The part can be cloned into a higher order level 1 composite clone using BsaI.