Parameters affecting the activity of antisense RNA sequences in tobacco protoplasts

François Guerineau, Robbie Waugh, John W.S. Brown

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Plasmids containing various fragments of the β-glucuronidase (GUS) gene were placed in antisense orientation downstream of the cauliflower mosaic virus 35S promoter and cotransfected with a 35S-gus construct into tobacco mesophyll protoplasts. None of the partial-length sequences were as effective as the full-length sequence in reducing GUS activity. The presence of a polyadenylation sequence downstream of the antisense sequence had an enhancing effect. The activity of the antisense sequence was largely affected by the incubation temperature of the transfected protoplasts. The chloramphenicol acetyltransferase (CAT) gene was fused to the gus coding sequence. When this construct was cotransfected with an antisense sequence directed against CAT, GUS activity was reduced. The implications of these results for the design and uses of antisense sequences are discussed.

Original languageEnglish
Pages (from-to)703-708
Number of pages6
JournalPlant Cell Reports
Volume13
Issue number12
DOIs
Publication statusPublished - 1 Sep 1994

Keywords

  • Antisense RNA
  • Gene expression
  • Protoplast
  • Reporter gene

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