Next generation synthetic vectors for transformation of the plastid genome of higher plants

Sugey Ramona Sinagawa-García, Tarinee Tungsuchat-Huang, Octavio Paredes-López, Pal Maliga

Research output: Contribution to journalArticlepeer-review

19 Scopus citations

Abstract

Plastid transformation vectors are E. coli plasmids carrying a plastid marker gene for selection, adjacent cloning sites and flanking plastid DNA to target insertions in the plastid genome by homologous recombination. We report here on a family of next generation plastid vectors carrying synthetic DNA vector arms targeting insertions in the rbcL-accD intergenic region of the tobacco (Nicotiana tabacum) plastid genome. The pSS22 plasmid carries only synthetic vector arms from which the undesirable restriction sites have been removed by point mutations. The pSS24 vector carries a c-Myc tagged spectinomycin resistance (aadA) marker gene whereas in vector pSS30 aadA is flanked with loxP sequences for post-transformation marker excision. The synthetic vectors will enable direct manipulation of passenger genes in the transformation vector targeting insertions in the rbcL-accD intergenic region that contains many commonly used restriction sites.

Original languageEnglish (US)
Pages (from-to)487-498
Number of pages12
JournalPlant Molecular Biology
Volume70
Issue number5
DOIs
StatePublished - Jul 1 2009

All Science Journal Classification (ASJC) codes

  • Agronomy and Crop Science
  • Genetics
  • Plant Science

Keywords

  • Homologous recombination
  • Nicotiana tabacum
  • Plastid transformation
  • Tobacco
  • Vector

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