Production of engineered minichromosome vectors via the introduction of telomere sequences

Nathaniel Graham, Nathan Swyers, Jon Cody, Morgan McCaw, Changzeng Zhao, James A. Birchler

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Artificial minichromosomes are non-integrating vectors capable of stably maintaining transgenes outside of the main chromosome set. The production of minichromosomes relies on telomere-mediated chromosomal truncation, which involves introducing transgenes and telomere sequences concurrently to the cell to truncate an endogenous chromosomal target. Two methods can be utilized; either the telomere sequences can be incorporated into a binary vector for transformation with Agrobacterium tumefaciens, or the telomere sequences can be co-introduced with transgenes during particle bombardment. In this protocol, the methods required to isolate and introduce telomere sequences are presented. Following the methods presented, standard transformation procedures can be followed to produce minichromosome containing plants.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
DOIs
StatePublished - Jan 1 2016

Publication series

NameMethods in Molecular Biology
Volume1469
ISSN (Print)1064-3745

Keywords

  • Agrobacterium tumefaciens transformation
  • Artificial chromosome
  • Biolistic bombardment
  • Chromosomal truncation
  • In-gel ligation
  • Minichromosome
  • Plant transformation
  • Telomere

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  • Cite this

    Graham, N., Swyers, N., Cody, J., McCaw, M., Zhao, C., & Birchler, J. A. (2016). Production of engineered minichromosome vectors via the introduction of telomere sequences. In Methods in Molecular Biology (Methods in Molecular Biology; Vol. 1469). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-4931-1_1