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.
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References
Gaeta RT, Masonbrink RE, Krishnaswamy L, Zhao C, Birchler JA (2012) Synthetic chromosome platforms in plants. Annu Rev Plant Biol 63:307–330
Yu W, Han F, Birchler JA (2007) Engineered minichromosomes in plants. Curr Opin Biotechnol 18:425–431
Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517
Ijdo JW, Wells RA, Baldini A, Reeders ST (1991) Improved telomere detection using a telomere repeat probe (TTAGGG)n generated by PCR. Nucleic Acids Res 19:4780
Green MR, Sambrook J (2012) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New York
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New York
Gaeta R, Masonbrink R, Zhao C, Sanyal A, Krishnaswamy L, Birchler J (2013) In vivo modification of a maize engineered minichromosome. Chromosoma 122:221–232
Yu W, Lamb J, Han F, Birchler J (2006) Telomere-mediated chromosomal truncation in maize. Proc Natl Acad Sci U S A 103:17331–17336
Lamb JC, Danilova T, Bauer MJ, Meyer UM, Holland JJ, Jensen MD, Birchler JA (2007) Single-gene detection and karyotyping using small-target fluorescence in situ hybridization on maize somatic chromosomes. Genetics 175:1047–1058
Acknowledgment
This work was supported by NSF grant IOS-1339198 from the Plant Genome Program.
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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: Murata, M. (eds) Chromosome and Genomic Engineering in Plants. Methods in Molecular Biology, vol 1469. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-4931-1_1
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DOI: https://doi.org/10.1007/978-1-4939-4931-1_1
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-4929-8
Online ISBN: 978-1-4939-4931-1
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