Team:Cambridge-JIC/Guide/Constructs/Reporters
From 2014.igem.org
Contents |
Reporters
Fluorescent proteins
The following fluorescent proteins have been used with consistent results in Marchantia: [http://parts.igem.org/Part:BBa_K165005 Venus YFP], MRFP1 and eGFP.
Localisation Tags
A range of localisation tags can be added to enable to distinguish sources of expression and to visualise individual cells and nuclei. These include: [http://parts.igem.org/Part:BBa_K1484104 N7] and [http://parts.igem.org/Part:BBa_K1484106 LTI].
These can be added instead of the stop codon for the reporters. They are incredibly useful when trying to distinguish individual cells, or different fluorescence channels.
These are clearly visible under fluorescence/confocal microscopes. Sample images include:
LTI:
N7:
Fluorescent transformants are visible first from 3-5 days after transformation.
Other reporters
GUS staining has been successfully used to characterise expression, as seen in in this [http://www.researchgate.net/publication/256611354_Comparison_of_the_MpEF1_and_CaMV35_promoters_for_application_in_Marchantia_polymorpha_overexpression_studies paper]