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 language||English (US)|
|Title of host publication||Methods in Molecular Biology|
|Publisher||Humana Press Inc.|
|State||Published - 2016|
|Name||Methods in Molecular Biology|
Bibliographical noteFunding Information:
This work was supported by NSF grant IOS-1339198 from the Plant Genome Program.
© Springer Science+Business Media New York 2016.
- Agrobacterium tumefaciens transformation
- Artificial chromosome
- Biolistic bombardment
- Chromosomal truncation
- In-gel ligation
- Plant transformation