Title: Exploration on Transformation of Saccharomy Cescerevisiae
Abstract:Electroporation and transformation induced by PEG/LiAc(polyethylene glycol/lithium acetate)is widely used for the transformation of Saccharomy cescerevisiae.However,it can not meet the highly efficien...Electroporation and transformation induced by PEG/LiAc(polyethylene glycol/lithium acetate)is widely used for the transformation of Saccharomy cescerevisiae.However,it can not meet the highly efficient transformation requirement(more than 105)of yeast-based pool screening.Based on the existing yeast transformation,the effects of different reagents and/or conditions on the transformation efficiency were further studied.The results show that the optimum period of Saccharomy cescerevisiae for transformation is the early stage of cell growth(OD600=0.6).In electroporation of Saccharomy cescerevisiae,the optimum concentration of DTT is 120 mmol/L,and that of LiAc is 300 mmol/L.Combination of DTT and LiAc will greatly promote the transformation efficiency to 7.15×105 cfu/μg.In transformation induced by PEG/LiAc,the transformation efficiency is increased to 2.75×105 cfu/μg when the concentration of LiAc is 300 mmol/L,which is much higher than the efficiency reported by other researchers.At the same time,it is confirmed that the optimum method is very useful for promoting the transformation efficiency of homologous recombination.This new optimized transformation system facilitates the research related to saccharomy cescerevisiae.Read More
Publication Year: 2012
Publication Date: 2012-01-01
Language: en
Type: article
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