A general method for polyethylene-glycol-induced genetic transformation of bacteria and yeast

Robert J. Klebe; June V. Harriss; Z. Dave Sharp; Michael G. Douglas

doi:10.1016/0378-1119(83)90238-X

A general method for polyethylene-glycol-induced genetic transformation of bacteria and yeast

Robert J. Klebe, June V. Harriss, Z. Dave Sharp, Michael G. Douglas

Research output: Contribution to journal › Article › peer-review

372 Scopus citations

Abstract

Polyethylene glycol (PEG) can induce genetic transformation in both bacteria (Escherichia coli) and yeast (Saccharomyces cerevisiae) without cell wall removal. PEG-mediated transformation of E. coli is technically simple and yields transformants with an efficiency of 10⁶-10⁷ transformants/μg DNA. Detailed analysis of the parameters involved in PEG-mediated transformation of E. coli reveals basic differences between the PEG and standard CaCl₂ methods for transformation of E. coli. PEG-mediated transformation of yeast is far simpler than existing protoplast methods and is comparable in efficiency. The new methods described here for PEG-mediated genetic transformation may prove to be of general utility in performing genetic transformation in a wide variety of organisms.

Original language	English (US)
Pages (from-to)	333-341
Number of pages	9
Journal	Gene
Volume	25
Issue number	2-3
DOIs	https://doi.org/10.1016/0378-1119(83)90238-X
State	Published - Nov 1983

Keywords

CaCl
Escherichia coli
Saccharomyces cerevisiae
Transformation
polyethylene glycol
protoplasts
recombinant DNA

ASJC Scopus subject areas

Genetics

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10.1016/0378-1119(83)90238-X

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@article{14c70a668795467883bf765dccf5ff97,

title = "A general method for polyethylene-glycol-induced genetic transformation of bacteria and yeast",

abstract = "Polyethylene glycol (PEG) can induce genetic transformation in both bacteria (Escherichia coli) and yeast (Saccharomyces cerevisiae) without cell wall removal. PEG-mediated transformation of E. coli is technically simple and yields transformants with an efficiency of 106-107 transformants/μg DNA. Detailed analysis of the parameters involved in PEG-mediated transformation of E. coli reveals basic differences between the PEG and standard CaCl2 methods for transformation of E. coli. PEG-mediated transformation of yeast is far simpler than existing protoplast methods and is comparable in efficiency. The new methods described here for PEG-mediated genetic transformation may prove to be of general utility in performing genetic transformation in a wide variety of organisms.",

keywords = "CaCl, Escherichia coli, Saccharomyces cerevisiae, Transformation, polyethylene glycol, protoplasts, recombinant DNA",

author = "Klebe, {Robert J.} and Harriss, {June V.} and Sharp, {Z. Dave} and Douglas, {Michael G.}",

note = "Funding Information: Our thanks to Dr. Barbara Bachmann and the E. coli Genetic Stock Center for the gift of certain strains used in this study. We wish to thank Ms. Sharon Ray for aid in preparation of the manuscript. This study was supported, in part, by grants from the National March of Dimes, the National Cancer Institute (CA33074), and the National Science Foundation (PCM-8218137).",

year = "1983",

month = nov,

doi = "10.1016/0378-1119(83)90238-X",

language = "English (US)",

volume = "25",

pages = "333--341",

journal = "Gene",

issn = "0378-1119",

publisher = "Elsevier",

number = "2-3",

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T1 - A general method for polyethylene-glycol-induced genetic transformation of bacteria and yeast

AU - Klebe, Robert J.

AU - Harriss, June V.

AU - Sharp, Z. Dave

AU - Douglas, Michael G.

N1 - Funding Information: Our thanks to Dr. Barbara Bachmann and the E. coli Genetic Stock Center for the gift of certain strains used in this study. We wish to thank Ms. Sharon Ray for aid in preparation of the manuscript. This study was supported, in part, by grants from the National March of Dimes, the National Cancer Institute (CA33074), and the National Science Foundation (PCM-8218137).

PY - 1983/11

Y1 - 1983/11

N2 - Polyethylene glycol (PEG) can induce genetic transformation in both bacteria (Escherichia coli) and yeast (Saccharomyces cerevisiae) without cell wall removal. PEG-mediated transformation of E. coli is technically simple and yields transformants with an efficiency of 106-107 transformants/μg DNA. Detailed analysis of the parameters involved in PEG-mediated transformation of E. coli reveals basic differences between the PEG and standard CaCl2 methods for transformation of E. coli. PEG-mediated transformation of yeast is far simpler than existing protoplast methods and is comparable in efficiency. The new methods described here for PEG-mediated genetic transformation may prove to be of general utility in performing genetic transformation in a wide variety of organisms.

AB - Polyethylene glycol (PEG) can induce genetic transformation in both bacteria (Escherichia coli) and yeast (Saccharomyces cerevisiae) without cell wall removal. PEG-mediated transformation of E. coli is technically simple and yields transformants with an efficiency of 106-107 transformants/μg DNA. Detailed analysis of the parameters involved in PEG-mediated transformation of E. coli reveals basic differences between the PEG and standard CaCl2 methods for transformation of E. coli. PEG-mediated transformation of yeast is far simpler than existing protoplast methods and is comparable in efficiency. The new methods described here for PEG-mediated genetic transformation may prove to be of general utility in performing genetic transformation in a wide variety of organisms.

KW - CaCl

KW - Escherichia coli

KW - Saccharomyces cerevisiae

KW - Transformation

KW - polyethylene glycol

KW - protoplasts

KW - recombinant DNA

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U2 - 10.1016/0378-1119(83)90238-X

DO - 10.1016/0378-1119(83)90238-X

M3 - Article

C2 - 6363214

AN - SCOPUS:0021071824

SN - 0378-1119

VL - 25

SP - 333

EP - 341

JO - Gene

JF - Gene

IS - 2-3

ER -

A general method for polyethylene-glycol-induced genetic transformation of bacteria and yeast

Abstract

Keywords

ASJC Scopus subject areas

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