HIGH-EFFICIENCY TRANSFORMATION OF INTACT YEAST-CELLS BY ELECTRIC-FIELD PULSES

被引：147

作者：

MEILHOC, E

MASSON, JM

TEISSIE, J

机构：

[1] INST NATL SCI APPL,CTR TRANSFERT BIOTECHNOL MICROBIOL,CNRS,UA 544,AVE RANGUEIL,F-31077 TOULOUSE,FRANCE

[2] CNRS,CTR RECH BIOCHIM & GENET CELLULAIRES,UPR 27,F-31062 TOULOUSE,FRANCE

来源：

BIO-TECHNOLOGY | 1990年 / 8卷 / 03期

关键词：

D O I：

10.1038/nbt0390-223

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

We have developed an efficient method that electrically introduces DNA into intact yeast cells. Saccharomyces cerevisiae was used as a model in order to optimize the transformation protocol. Transformation efficiencies of 107 transformation of plasmid DNA were obtained with a square wave electric pulse of 2.7 kV/cm during 15 milliseconds. The technique is simple and rapid. Even small quantities of DNA (100 pg) can be used to transform 108 cells. Important parameters are the pulse field strength and duration. Pretreatment of the yeast cells in the early phase of exponential growth with dithiothreitol increases transformation efficiency. The method has been successfully applied to various strains of 5. cerevisiae as well as to other types of yeast. © 1990 Nature Publishing Group.

引用

页码：223 / 227

页数：5

共 34 条

[1]

ANDREASON GL, 1988, BIOTECHNIQUES, V6, P650

[2]

AUSUBEL FM, 1989, CURRENT PROTOCOL MOL, V2

[3] TRANSFORMATION OF YEAST BY A REPLICATING HYBRID PLASMID [J].