Gene Transfer to Yeast with New Vacuum and ...Vacuum and Electroporation Method) Gene Transfer to...
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Gene Transfer to Yeast (Vacuum and Electroporation Method) Gene Transfer to Yeast with New Vacuum and Electroporation Method (1/3)
Transcript of Gene Transfer to Yeast with New Vacuum and ...Vacuum and Electroporation Method) Gene Transfer to...
Gene Transfer to Yeast(Vacuum and Electroporation Method)
Gene Transfer to Yeastwith
New Vacuum and Electroporation Method
(1/3)
(2/4)
Specific name Saccharomyces
cerevisiae
Strain BY4700(Sequenced all its genome)
Genotype MATa ura3Δ0Ura3 / Orotidine-5‘-phosphate decarboxylase missing
Department of Applied Chemistry and Chemical Engineering,Faculty of Engineering,Yamaguchi University
Fig.1 Procedure for Gene Transfer to Yeast
Gene Transfer to YeastGene Transfer Procedure
Vector DNA
Amylase gene
Uracil synthesis
gene
Host YeastYeast that can not synthesize uracil
Unable to culture in urasil missing medium (minimum medium)
Gene Transfer
Select transferred gene in minimum medium
Minimum medium
Amylase gene
Uracil synthesis
gene
recombinant
Recombinant secretes mold amylase
Amylolysis part by amylase
Starch iodine reaction
Starch medium
(3/4)
Transfectant Wild type
Gene Transfer to YeastCultivation of Yeast
(4/4)
HS : Heat ShockV: VacuumEP : ElectroporationV + EP : Vacuum + Electroporation
Fig.1 Efficiency of Yeast Transformation
V+EP is best
コロニー数
0
50
100
150
200
HS V EP V+EP
コロニー数
A. Heat Shock Method （conventionally）
B. Vacuum + Electroporation
Gene Transfer to YeastEfficiency of Yeast Transformation
No. of Colony
