The pCLEAN dual binary vector system for Agrobacterium-mediated plant transformation

Plant Physiol. 2007 Dec;145(4):1211-9. doi: 10.1104/pp.107.108563. Epub 2007 Oct 11.

Abstract

The development of novel transformation vectors is essential to the improvement of plant transformation technologies. Here, we report the construction and testing of a new multifunctional dual binary vector system, pCLEAN, for Agrobacterium-mediated plant transformation. The pCLEAN vectors are based on the widely used pGreen/pSoup system and the pCLEAN-G/pCLEAN-S plasmids are fully compatible with the existing pGreen/pSoup vectors. A single Agrobacterium can harbor (1) pCLEAN-G and pSoup, (2) pGreen and pCLEAN-S, or (3) pCLEAN-G and pCLEAN-S vector combination. pCLEAN vectors have been designed to enable the delivery of multiple transgenes from distinct T-DNAs and/or vector backbone sequences while minimizing the insertion of superfluous DNA sequences into the plant nuclear genome as well as facilitating the production of marker-free plants. pCLEAN vectors contain a minimal T-DNA (102 nucleotides) consisting of direct border repeats surrounding a 52-nucleotide-long multiple cloning site, an optimized left-border sequence, a double left-border sequence, restriction sites outside the borders, and two independent T-DNAs. In addition, selectable and/or reporter genes have been inserted into the vector backbone sequence to allow either the counter-screening of backbone transfer or its exploitation for the production of marker-free plants. The efficiency of the different pCLEAN vectors has been assessed using transient and stable transformation assays in Nicotiana benthamiana and/or Oryza sativa.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA, Bacterial
  • Genetic Engineering*
  • Genetic Vectors*
  • Genome, Plant
  • Molecular Sequence Data
  • Oryza / genetics*
  • Oryza / microbiology
  • Plasmids*
  • Rhizobium / genetics*
  • Tobacco / genetics*
  • Tobacco / microbiology
  • Transformation, Genetic*
  • Transgenes

Substances

  • DNA, Bacterial
  • T-DNA

Associated data

  • GENBANK/EU186081
  • GENBANK/EU186082
  • GENBANK/EU186083
  • GENBANK/EU186084
  • GENBANK/EU186085
  • GENBANK/EU186086
  • GENBANK/EU186087
  • GENBANK/EU186088
  • GENBANK/EU186089
  • GENBANK/EU186090
  • GENBANK/EU186091
  • GENBANK/EU186092