An intronless form of the tobacco extensin gene terminator strongly enhances transient gene expression in plant leaves

Sun Hee Rosenthal, Andrew G. Diamos, Hugh Mason

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Key message: We have found interesting features of a plant gene (extensin) 3′ flanking region, including extremely efficient polyadenylation which greatly improves transient expression of transgenes when an intron is removed. Its use will greatly benefit studies of gene expression in plants, research in molecular biology, and applications for recombinant proteins. Abstract: Plants are a promising platform for the production of recombinant proteins. To express high-value proteins in plants efficiently, the optimization of expression cassettes using appropriate regulatory sequences is critical. Here, we characterize the activity of the tobacco extensin (Ext) gene terminator by transient expression in Nicotiana benthamiana, tobacco, and lettuce. Ext is a member of the hydroxyproline-rich glycoprotein (HRGP) superfamily and constitutes the major protein component of cell walls. The present study demonstrates that the Ext terminator with its native intron removed increased transient gene expression up to 13.5-fold compared to previously established terminators. The enhanced transgene expression was correlated with increased mRNA accumulation and reduced levels of read-through transcripts, which could impair gene expression. Analysis of transcript 3′-ends found that the majority of polyadenylated transcripts were cleaved at a YA dinucleotide downstream from a canonical AAUAAA motif and a UG-rich region, both of which were found to be highly conserved among related extensin terminators. Deletion of either of these regions eliminated most of the activity of the terminator. Additionally, a 45 nt polypurine sequence ~ 175 nt upstream from the polyadenylation sites was found to also be necessary for the enhanced expression. We conclude that the use of Ext terminator has great potential to benefit the production of recombinant proteins in plants.

Original languageEnglish (US)
Pages (from-to)1-15
Number of pages15
JournalPlant Molecular Biology
DOIs
StateAccepted/In press - Feb 10 2018

Fingerprint

extensin
Plant Leaves
Recombinant Proteins
Tobacco
Polyadenylation
tobacco
Transgenes
Gene Expression
gene expression
Introns
recombinant proteins
3' Flanking Region
Genes
leaves
Plant Genes
Lettuce
Plant Proteins
genes
Hydroxyproline
Cell Wall

Keywords

  • Intron
  • Plant based expression
  • Polyadenylation
  • Polypurine sequence
  • Read-through transcription
  • Tobacco extensin terminator

ASJC Scopus subject areas

  • Agronomy and Crop Science
  • Genetics
  • Plant Science

Cite this

An intronless form of the tobacco extensin gene terminator strongly enhances transient gene expression in plant leaves. / Rosenthal, Sun Hee; Diamos, Andrew G.; Mason, Hugh.

In: Plant Molecular Biology, 10.02.2018, p. 1-15.

Research output: Contribution to journalArticle

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