title: Production of Engineered Minichromosome Vectors via the Introduction of Telomere Sequences.
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.
authors.affiliation: Division of Biological Sciences, University of Missouri, Columbia, MO, 65211, USA.
authors.forename: Nathaniel
authors.initials: N
authors.lastname: Graham
authors.affiliation: Division of Biological Sciences, University of Missouri, Columbia, MO, 65211, USA.
authors.forename: Nathan
authors.initials: N
authors.lastname: Swyers
authors.affiliation: Division of Biological Sciences, University of Missouri, Columbia, MO, 65211, USA.
authors.forename: Jon
authors.initials: J
authors.lastname: Cody
authors.affiliation: Division of Biological Sciences, University of Missouri, Columbia, MO, 65211, USA.
authors.forename: Morgan
authors.initials: M
authors.lastname: McCaw
authors.affiliation: Division of Biological Sciences, University of Missouri, Columbia, MO, 65211, USA.
authors.forename: Changzeng
authors.initials: C
authors.lastname: Zhao
authors.affiliation: Division of Biological Sciences, University of Missouri, Columbia, MO, 65211, USA. BirchlerJ@Missouri.edu.
authors.forename: James A
authors.initials: JA
authors.lastname: Birchler
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics; D022201:Chromosomes, Artificial; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D016615:Telomere; D019076:Transgenes
keywords: Agrobacterium tumefaciens transformation; Artificial chromosome; Biolistic bombardment; Chromosomal truncation; In-gel ligation; Minichromosome; Plant transformation; Telomere