PARAMETERS AFFECTING LITHIUM-ACETATE MEDIATED TRANSFORMATION OF SACCHAROMYCES-CEREVISIAE AND DEVELOPMENT OF A RAPID AND SIMPLIFIED PROCEDURE

被引:134
作者
SONI, R [1 ]
CARMICHAEL, JP [1 ]
MURRAY, JAH [1 ]
机构
[1] UNIV CAMBRIDGE,INST BIOTECHNOL,TENNIS COURT RD,CAMBRIDGE CB2 1QT,ENGLAND
关键词
YEAST; SACCHAROMYCES-CEREVISIAE; TRANSFORMATION; PLASMID;
D O I
10.1007/BF00351857
中图分类号
Q3 [遗传学];
学科分类号
071007 [遗传学]; 090102 [作物遗传育种];
摘要
We have compared a number of procedures for the transformation of whole cells of the yeast Saccharomyces cerevisiae and assessed the effects of dimethylsulphoxide (DMSO) or ethanol, both of which have been reported to enhance transformation efficiency. We find that simplified methods benefit from the addition of one of these compounds, and although differences are observed between strains as to the more beneficial reagent, peak transformation efficiency is, in general, obtained with 10% DMSO or 10% EtOH. Increases of between six- and 50-fold are observed, despite a reduction in cell viability, and at this concentration the two compounds are not additive in their effects. The optimum level appears to depend on a balance between improved DNA uptake and reduced cell viability. As a result of this work we present a straightforward and rapid transformation procedure.
引用
收藏
页码:455 / 459
页数:5
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