Part:BBa_K530008:Experience

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Team Kyoto 2017

S. cerevisiae expressing GFP from this TDH3 promoter was cultured and observed with a fluorescence microscope.
The strong green fluorescence was observed. (fig 1 )

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

B. xylophilus was cultured using this green fluorescent yeast as a bait. After several days, confocal observation confirmed that the intestine of B. xylophilus was filled with EGFP. (fig 2 )

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

This means that using the TDH3 promoter (BBa_K530008) makes it possible to express a very large amount of EGFP in S. cerevisiae, and that its expression level is sufficient to trace the intestinal tract of nematodes feeding on yeast.
The above results indicate that BBa_K530008 and BBa_K1875003 work effectively as a new usage that has not been attempted so far, greatly extending the function of each part.

User Reviews

UNIQc81e3ad0b012b89c-partinfo-00000001-QINU UNIQc81e3ad0b012b89c-partinfo-00000002-QINU

NAU-CHINA 2017 iGEM

Figure. Fluorescence intensity of different colonies of TDH3 promoter and improved TDH3 promoter.

We construct the device： TDH3 promoter+GFP+ADH1 terminator TDH3 promoter is a truncated promoter, GFP is a mutant. These is a invariable sequence upstream of GFP coding sequence.

The responses of promoter strengths of artificial TDH3 promoter and native promoter TDH3 were comprehensively compared. The strength of improved TDH3 was always higher than native promoter TDH3，but varied along with the genetic background of host.

TDH3 from kit plate

TDH3 from kit plate,the construct was transfected into Saccharomyces cerevisiae cells, the green fluorescence was observed under fluorescence microscope clearly. It indicates this part could work.