Difference between revisions of "Part:BBa K105026"

Revision as of 11:59, 17 September 2015

Gal1 promoter

This is an aquivalent to J63006. We reconstructed this BioBrick because the length of the part we received from the registery didn't correspond to the J63006. Neither the sequence from the MIT sequencing did.

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Sequence and Features

Application

Group: FAFU-CHINA

Author: Ruicheng Dai & Changlong Lu

Summary:The effect of GAL1 promoter induced by galactose

galactose inductive effect In order to improve the prokaryotic system , we are going to introduce the prokaryotic system into yeast (eukaryotic systems). However, considering there is no T7 RNAP gene in yeast, we will design T7 RNAP-pYES2 plasmid to express T7 RNAP. Meanwhile, because we do not know the effect of GAL1 promoter induced by different concentrations of galactose, we use pyes2 plasmid to express GFP protein firstly. Through measuring the fluorescence value under the induction of different concentrations of galactose and at different time intervals, we try to determine the GAL1 induced effect.

The inductive effect of different concentration galactose We transformed GFP- pYES2 into the yeast and let the recombinant strain grow to mid-log phase in YEPD medium containing 2% glucose. And then we transferred it to medium, containing 2%, 0.1% or 0.02% galactose, and assayed for GFP fluorescence.

The inductive effect of different time length The yeast transformants were grown in YEPD medium. At a cell density of 2 ×107 ml-1,cultures were washed and galactose added to a final concentration of 2% and the fluorescence value of samples was measured every 5h.