Part:BBa_K5160005
mCherry
Overview
As a red fluorescent protein, we selected part of its sequence as a control for monitoring the effectiveness of TRV viral vectors.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design
When carrying out the design, considering that we need to set up a control group to avoid the influence of the virus itself on the experiment, we purposely constructed the mCherry protein on the TRV2_35S promoter_mCherry_NOS terminator.
Source
mCherry is derived from mushroom coral and has been artificially modified to stabilize fluorescent proteins.
Biology
mCherry is a red fluorescent protein from the mushroom coral that is commonly used to label and trace certain molecules and cellular components. The benefits of mCherry over other fluorescence are that it can be co-labeled with the most used green fluorescent protein (GFP) and that mCherry has better photostability than other monomeric fluorescent proteins. In our project, mCherry was used for TRV virus-mediated transient expression. In this regard, intact mCherry has the following main roles:
1、As a visualization marker mCherry is capable of emitting red fluorescence, which can be used to visualize the process of virus infestation and the extent of spread in experiments. Through the fluorescence microscope and other equipment, it can track the spreading path of the virus in plant tissues, helping researchers to determine the site and speed of virus infestation.
2、Monitoring the effectiveness of viral vectors When mCherry is combined with the TRV virus vector, the fluorescence signal of mCherry can be detected to determine whether the virus vector has successfully infected the plant cells. If the fluorescence of mCherry is observed in plant tissues, it means that the viral vector has entered the cells and started to express fluorescent proteins, thus indirectly proving the effectiveness of the viral vector.
3、Assist in the study of gene silencing effect In some gene silencing experiments based on TRV virus, mCherry can be constructed into viral vectors together with the target gene. By observing the change of fluorescence intensity of mCherry, we can initially determine whether the target gene has been successfully silenced. If the target gene is silenced, it may affect the physiological processes related to the gene, which in turn leads to changes in the expression level of mCherry.
In our experiments, we intercepted part of the sequence of mCherry as a control. Part of the mCherry does not have fluorescent properties and is only used for comparison with the experimental group.
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