Transient expression of genes in the oomycete Phytophthora infestans using Bremia lactucae regulatory sequences

Howard S. Judelson, Richard W Michelmore

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Vectors containing fusions between the reporter gene β-glucuronidase (GUS) and transcriptional regulatory signals from the ham34 and hsp70 genes of the oomycete pathogen, Bremia lactucae, were introduced into protoplasts of the related fungus Phytophthora infestans. Transient expression of the GUS gene was detected when DNA was introduced into protoplasts of P. infestans by treatments with polyethylene glycol and CaCl2 with cationic liposomes, or by electroporation. After optimization of each procedure, using the transient expression assays, cationic liposomes were identified as the superior method for DNA uptake. Vectors containing the 5′hsp70 sequences and 3′ham34 sequences resulted in the maximum level of GUS activity. The identification of functional vectors for transformation, and of optimal methods for introducing DNA, should assist in achieving stable transformation of P. infestans and other oomycete the fungi.

Original languageEnglish (US)
Pages (from-to)453-459
Number of pages7
JournalCurrent Genetics
Volume19
Issue number6
DOIs
StatePublished - Jun 1991

Fingerprint

Phytophthora infestans
Oomycetes
Glucuronidase
Protoplasts
Gene Expression
Liposomes
DNA
Fungi
Electroporation
Reporter Genes
Genes
Therapeutics

Keywords

  • β-glucuronidase
  • Fungi
  • Late blight
  • Transformation

ASJC Scopus subject areas

  • Genetics

Cite this

Transient expression of genes in the oomycete Phytophthora infestans using Bremia lactucae regulatory sequences. / Judelson, Howard S.; Michelmore, Richard W.

In: Current Genetics, Vol. 19, No. 6, 06.1991, p. 453-459.

Research output: Contribution to journalArticle

Judelson, Howard S. ; Michelmore, Richard W. / Transient expression of genes in the oomycete Phytophthora infestans using Bremia lactucae regulatory sequences. In: Current Genetics. 1991 ; Vol. 19, No. 6. pp. 453-459.
@article{d7672f8067f34e0cbae0d3b1e060d1f3,
title = "Transient expression of genes in the oomycete Phytophthora infestans using Bremia lactucae regulatory sequences",
abstract = "Vectors containing fusions between the reporter gene β-glucuronidase (GUS) and transcriptional regulatory signals from the ham34 and hsp70 genes of the oomycete pathogen, Bremia lactucae, were introduced into protoplasts of the related fungus Phytophthora infestans. Transient expression of the GUS gene was detected when DNA was introduced into protoplasts of P. infestans by treatments with polyethylene glycol and CaCl2 with cationic liposomes, or by electroporation. After optimization of each procedure, using the transient expression assays, cationic liposomes were identified as the superior method for DNA uptake. Vectors containing the 5′hsp70 sequences and 3′ham34 sequences resulted in the maximum level of GUS activity. The identification of functional vectors for transformation, and of optimal methods for introducing DNA, should assist in achieving stable transformation of P. infestans and other oomycete the fungi.",
keywords = "β-glucuronidase, Fungi, Late blight, Transformation",
author = "Judelson, {Howard S.} and Michelmore, {Richard W}",
year = "1991",
month = "6",
doi = "10.1007/BF00312736",
language = "English (US)",
volume = "19",
pages = "453--459",
journal = "Current Genetics",
issn = "0172-8083",
publisher = "Springer Verlag",
number = "6",

}

TY - JOUR

T1 - Transient expression of genes in the oomycete Phytophthora infestans using Bremia lactucae regulatory sequences

AU - Judelson, Howard S.

AU - Michelmore, Richard W

PY - 1991/6

Y1 - 1991/6

N2 - Vectors containing fusions between the reporter gene β-glucuronidase (GUS) and transcriptional regulatory signals from the ham34 and hsp70 genes of the oomycete pathogen, Bremia lactucae, were introduced into protoplasts of the related fungus Phytophthora infestans. Transient expression of the GUS gene was detected when DNA was introduced into protoplasts of P. infestans by treatments with polyethylene glycol and CaCl2 with cationic liposomes, or by electroporation. After optimization of each procedure, using the transient expression assays, cationic liposomes were identified as the superior method for DNA uptake. Vectors containing the 5′hsp70 sequences and 3′ham34 sequences resulted in the maximum level of GUS activity. The identification of functional vectors for transformation, and of optimal methods for introducing DNA, should assist in achieving stable transformation of P. infestans and other oomycete the fungi.

AB - Vectors containing fusions between the reporter gene β-glucuronidase (GUS) and transcriptional regulatory signals from the ham34 and hsp70 genes of the oomycete pathogen, Bremia lactucae, were introduced into protoplasts of the related fungus Phytophthora infestans. Transient expression of the GUS gene was detected when DNA was introduced into protoplasts of P. infestans by treatments with polyethylene glycol and CaCl2 with cationic liposomes, or by electroporation. After optimization of each procedure, using the transient expression assays, cationic liposomes were identified as the superior method for DNA uptake. Vectors containing the 5′hsp70 sequences and 3′ham34 sequences resulted in the maximum level of GUS activity. The identification of functional vectors for transformation, and of optimal methods for introducing DNA, should assist in achieving stable transformation of P. infestans and other oomycete the fungi.

KW - β-glucuronidase

KW - Fungi

KW - Late blight

KW - Transformation

UR - http://www.scopus.com/inward/record.url?scp=0025829427&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025829427&partnerID=8YFLogxK

U2 - 10.1007/BF00312736

DO - 10.1007/BF00312736

M3 - Article

AN - SCOPUS:0025829427

VL - 19

SP - 453

EP - 459

JO - Current Genetics

JF - Current Genetics

SN - 0172-8083

IS - 6

ER -