Difference between revisions of "Part:BBa K2403003"

Revision as of 11:02, 1 November 2017 (view source) TongYou (Talk | contribs) (→‎Characterization) ← Older edit		Revision as of 02:48, 2 November 2017 (view source) TongYou (Talk | contribs) (→‎Characterization) Newer edit →
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	<img src="https://static.igem.org/mediawiki/parts/8/82/B._xEGFP.jpg" alt="<i>B. xylophilus</i> filled with EGFP in its intestine" style="float:center;width:350px;height:250px;">		<img src="https://static.igem.org/mediawiki/parts/8/82/B._xEGFP.jpg" alt="<i>B. xylophilus</i> filled with EGFP in its intestine" style="float:center;width:350px;height:250px;">
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−	Figure2 : <i>B. xylophilus</i> full of EGFP in its esophagus.	+	Figure2 : <i>B. xylophilus</i> full of EGFP in its intestine.
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Revision as of 02:48, 2 November 2017

TDH3 promoter>EGFP protein

We expressed this gene in Saccharomyces cerevisiae and feed it to B.xylophilus.

Usage and Biology

Aiming for feeding RNAi of B. xylophilus, our team had to confirm that B. xylophilus surely eats S. cerevisiae. To confirm this, we expressed high levels of EGFP in S. cerevisiae by combining BBa_K530008(TDH3 promoter) and BBa_K1875003 (EGFP) and creating BBa_K2403003. Since this part produces a very strong green fluorescence, it is useful for tracking yeast.

Characterization

Our team observed that yeast fluoresces EGFP as shown in fig.1. We cultured nematodes using this green fluorescent yeast as a bait. A few days later, we collected and observed the nematodes with a confocal microscope.
It was confirmed that there were individuals emitting strong green fluorescence, and it was possible to confirm for the first time in the world that this nematode is eating S. cerevisiae (fig 2). This fluorescence illuminates the entire intestinal tract of B. xylophilus, and it is shown that this yeast is also useful as a tool for seeing the intestinal structure of nematodes.

Figure1: Observation of S. cerevisiae expressing EGFP by this TDH3 promoter with a fluorescence microscope.

Figure2 : B. xylophilus full of EGFP in its intestine.

According to the above results, this new part makes sufficiently high intensity EGFP expressed in yeast, and the amount is sufficient to observe the intestinal structure of the nematode feeding the yeast. Such usage greatly extends the use applications of BBa_K530008 and BBa_K1875003, making it possible to conduct similar experiments on other nematode species with stylets that prey on yeast.

Reference

BBa_K530008
BBa_K1875003

Sequence and Features