EBY100 yeast display system host strain酵母表面展示宿主菌

Cat# NTCC-EBY100

Yeast NTCC^TMEBY100 strain

NTCC^TMEBY100 expresses the AGA1 gene under control of the GAL1 promoter. It was created by integrating the vector pIU211 into the AGA1 locus of the Saccharomyces cerevisiae strain BJ5465 (MATa ura 3-52 trp 1 leu2Δ1 his3Δ200 pep4:HIS3 prb1Δ1.6R can1 GAL)(Boder and Wittrup, 1997). pIU211 contains the AGA1 gene regulated by the GAL promoter and a URA3 selectable marker. The vector was linearized for integration at AGA1 using BsiW I. BsiW I cuts in the AGA1 gene, ensuring that the construct integrates at the AGA1 locus.

To transform NTCC^TMEBY100 you will need the following reagents.

? Minimal dextrose plates containing leucine and tryptophan

? YPD medium

? Minimal dextrose plates containing leucine

? 20% Glucose

? 0.5-5 μg plasmid DNA (DNA may be isolated from E. coli using your method of

choice)

Transformation of NTCC^TMEBY100

Follow the general steps below to transform NTCC^TMEBY100.

1. Take the glycerol stock of NTCC^TMEBY100 and streak out on a minimal dextrose plate

containing leucine and tryptophan. Incubate at 30°C until colonies appear (1-2 days).

2. Use your method of choice to prepare competent cells and transform with pYD1 or

pYD1 containing your gene of interest.

3. Plate the transformation reaction (50-150 μl) on minimal dextrose plates containing

leucine. Incubate the plates at 30°C for 2 to 4 days until single colonies appear.

Once you have transformants, you are ready to test for display of your fusion protein.

Proceed to the next page to induce expression of your fusion protein.

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