Chimeric vector construction for higher-plant transformation

Ervin Balázs, Salah Bouzoubaa, Hubert Guilley, Gerard Jonard, Jerzy Paszkowski, Ken Richards

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

A chimeric vector pKR612B1 was developed containing the neomycin phosphotransferase (APH) gene from the Tn5 transposon under the control of the gene VI promoter of cauliflower mosaic virus (CaMV), and was used to transform higher plant protoplasts. Plasmid pDOB612, the parental vector of pKR612B1, has two unique restriction sites, SmaI and BamHI, positioned just downstream of the CaMV gene VI promoter sequence. These unique cloning sites can be used for any kind of gene insertion into this vector. Using the polyethylene glycol transformation procedure, a large number of turnip and tobacco protoplasts were transformed and proved to be resistant to kanamycin (Km). From tobacco protoplasts whole Km-resistant plants were regenerated and shown to contain the integrated foreign gene. APH activity was detected in both transformed calli and in regenerated plants. DNA from transformed clones was analysed by Southern blot hybridization, showing the presence of the Tn5-derived gene.

Original languageEnglish
Pages (from-to)343-348
Number of pages6
JournalGene
Volume40
Issue number2-3
DOIs
Publication statusPublished - 1985

Keywords

  • Recombinant DNA
  • callus
  • cauliflower mosaic virus promoter, Tn5
  • kanamycin resistance
  • plant protoplasts
  • transposon

ASJC Scopus subject areas

  • Genetics

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    Balázs, E., Bouzoubaa, S., Guilley, H., Jonard, G., Paszkowski, J., & Richards, K. (1985). Chimeric vector construction for higher-plant transformation. Gene, 40(2-3), 343-348. https://doi.org/10.1016/0378-1119(85)90059-9