A recombinase-mediated transcriptional induction system in transgenic plants

Plant Mol Biol. 2001 Jan;45(1):41-9. doi: 10.1023/a:1006402308365.

Abstract

We constructed and tested a Cre-loxP recombination-mediated vector system termed pCrox for use in transgenic plants. In this system, treatment of Arabidopsis under inducing conditions mediates an excision event that removes an intervening piece of DNA between a promoter and the gene to be expressed. The system developed here uses a heat-shock-inducible Cre to excise a DNA fragment flanked by lox sites, thereby generating a constitutive GUS reporter gene under control of the CaMV 35S promoter. Heat-shock-mediated excision of several, independent lines resulted in varying degrees of recombination-mediated GUS activation. Induction was shown to be possible at essentially any stage of plant growth. This single vector system circumvents the need for genetic crosses required by other, dual recombinase vector systems. The pCrox system may prove particularly useful in instances where transgene over-expression, or under-expression by antisense, would otherwise affect embryo, seed or seedling viability.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetyltransferases / genetics
  • Acetyltransferases / metabolism
  • Arabidopsis / genetics
  • Blotting, Southern
  • DNA, Plant / genetics
  • DNA, Plant / metabolism
  • Gene Expression Regulation, Plant
  • Genetic Vectors
  • Glucuronidase / genetics
  • Glucuronidase / metabolism
  • Hot Temperature
  • Integrases / genetics
  • Integrases / metabolism*
  • Plants, Genetically Modified / genetics*
  • Plasmids / genetics
  • Polymerase Chain Reaction
  • RNA, Plant / genetics
  • RNA, Plant / metabolism
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Transcriptional Activation*
  • Viral Proteins*

Substances

  • DNA, Plant
  • RNA, Plant
  • Recombinant Fusion Proteins
  • Viral Proteins
  • Acetyltransferases
  • phosphinothricin N-acetyltransferase
  • Cre recombinase
  • Integrases
  • Glucuronidase