The pFF plasmids

cassettes utilising CaMV sequences for expression of foreign genes in plants

Marja C.P. Timmermans, Pal Maliga, Jeffrey Vieira, Joachim Messing

Research output: Contribution to journalArticle

123 Citations (Scopus)

Abstract

A plant expression cassette was constructed using the cauliflower mosaic virus 35S 5′ regulatory region with the enhancer duplicated and the 35S polyadenylation signal. Insertion of a polylinker between the transcription initiation and polyadenylation sites allows for easy cloning of genes. To test the usefulness of the cassette chimeric bacterial genes were prepared. The constructs were introduced into Nicotiana tabacum suspension culture cells by the particle bombardment process. Expression of the β-glucuronidase reporter gene was verified by histochemical staining. Stable kanamycin and hygromycin resistant transgenic lines were obtained after introduction of chimeric genes encoding the enzymes neomycin phosphotransferase and hygromycin B phosphotransferase, respectively. The number of stable transformants was approximately 2% of the cells that transiently expressed the β-glucuronidase reporter gene.

Original languageEnglish (US)
Pages (from-to)333-344
Number of pages12
JournalJournal of Biotechnology
Volume14
Issue number3-4
DOIs
StatePublished - Jan 1 1990

Fingerprint

Plant Genes
Polyadenylation
hygromycin-B kinase
Glucuronidase
Reporter Genes
Plasmids
Framycetin
Genes
Hygromycin B
Kanamycin Kinase
Caulimovirus
Gene Expression
Bacterial Genes
Kanamycin
Nucleic Acid Regulatory Sequences
Transcription Initiation Site
Tobacco
Organism Cloning
Suspensions
Cell Culture Techniques

All Science Journal Classification (ASJC) codes

  • Applied Microbiology and Biotechnology
  • Bioengineering
  • Biotechnology

Keywords

  • CaMV sequence
  • Foreign gene
  • Plant
  • Plasmid, pFF

Cite this

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title = "The pFF plasmids: cassettes utilising CaMV sequences for expression of foreign genes in plants",
abstract = "A plant expression cassette was constructed using the cauliflower mosaic virus 35S 5′ regulatory region with the enhancer duplicated and the 35S polyadenylation signal. Insertion of a polylinker between the transcription initiation and polyadenylation sites allows for easy cloning of genes. To test the usefulness of the cassette chimeric bacterial genes were prepared. The constructs were introduced into Nicotiana tabacum suspension culture cells by the particle bombardment process. Expression of the β-glucuronidase reporter gene was verified by histochemical staining. Stable kanamycin and hygromycin resistant transgenic lines were obtained after introduction of chimeric genes encoding the enzymes neomycin phosphotransferase and hygromycin B phosphotransferase, respectively. The number of stable transformants was approximately 2{\%} of the cells that transiently expressed the β-glucuronidase reporter gene.",
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The pFF plasmids : cassettes utilising CaMV sequences for expression of foreign genes in plants. / Timmermans, Marja C.P.; Maliga, Pal; Vieira, Jeffrey; Messing, Joachim.

In: Journal of Biotechnology, Vol. 14, No. 3-4, 01.01.1990, p. 333-344.

Research output: Contribution to journalArticle

TY - JOUR

T1 - The pFF plasmids

T2 - cassettes utilising CaMV sequences for expression of foreign genes in plants

AU - Timmermans, Marja C.P.

AU - Maliga, Pal

AU - Vieira, Jeffrey

AU - Messing, Joachim

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Y1 - 1990/1/1

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AB - A plant expression cassette was constructed using the cauliflower mosaic virus 35S 5′ regulatory region with the enhancer duplicated and the 35S polyadenylation signal. Insertion of a polylinker between the transcription initiation and polyadenylation sites allows for easy cloning of genes. To test the usefulness of the cassette chimeric bacterial genes were prepared. The constructs were introduced into Nicotiana tabacum suspension culture cells by the particle bombardment process. Expression of the β-glucuronidase reporter gene was verified by histochemical staining. Stable kanamycin and hygromycin resistant transgenic lines were obtained after introduction of chimeric genes encoding the enzymes neomycin phosphotransferase and hygromycin B phosphotransferase, respectively. The number of stable transformants was approximately 2% of the cells that transiently expressed the β-glucuronidase reporter gene.

KW - CaMV sequence

KW - Foreign gene

KW - Plant

KW - Plasmid, pFF

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