Difference between revisions of "Part:BBa K1484104"

Revision as of 18:12, 10 October 2014

N7 nuclear localisation tag

Localisation tag for plant reporters, which localises them to the nucleus. Compatible with the Plant Syntax (RFC105) as a C-Terminal fusion.

Usage and Biology

Add this tag to the 3' of your ORF. IMPORTANT: this contains flanking restriction sites for use with RFC105 compatible assembly.

When used in conjunction with fluorescent reporters, nuclei are clearly visible with 5x magnification.

This tag (in Marchantia) does not appear to interfere with plant growth, in contrast to histone-binding localisation tags (for example, H2B), which may do so.

Its use is extensively recorded in the literature.

In Marchantia: [file:CAM14_gfpN7.tif] Image taken on a Leica D6000 confocal microscope, using excitation laser of 488nm and emission range 525-540.

This tag can be very useful for counting cells, finding cell seeds in order to use balloon segmentation (in conjunction with LTI) and also for visualizing multiple channels of fluorescent proteins across constructs.

In conjunction with image analysis techniques, the use of this tag can enable high-throughput quantification of gene expression [when standardized sample conditions are used].

As a simple example of the kind of analysis which can be performed with basic image analysis techniques:

The identification of nuclei was done using Python's scipy.measurements.image.ndlabel method. Sequence and Features