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EBY100酵母表面展示宿主菌 BioVector NTCC质粒载体菌种细胞基因保藏中心

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  • 货  号:EBY100酵母表面展示宿主菌
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EBY100酵母表面展示宿主菌


YeastEBY100 strain


EBY100expresses the AGA1 gene under control of the GAL1 promoter. It was created byintegrating the vector pIU211 into the AGA1 locus of the Saccharomycescerevisiae strain BJ5465 (MATa ura 3-52 trp 1 leu2Δ1 his3Δ200 pep4:HIS3 prb1Δ1.6Rcan1 GAL)(Boder and Wittrup, 1997). pIU211 contains the AGA1 gene regulated bythe GAL promoter and a URA3 selectable marker. The vector was linearized forintegration at AGA1 using BsiW I. BsiW I cuts in the AGA1 gene, ensuring thatthe construct integrates at the AGA1 locus.



Totransform EBY100 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 plasmidDNA (DNA may be isolated from E. coli using your method of

choice)



Transformationof EBY100

Follow the general steps below to transformEBY100.

1.Take the glycerol stock of EBY100 and streak out on a minimal dextrose plate

containingleucine 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

pYD1containing your gene of interest.

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

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

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

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


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