Graham Nathaniel, Swyers Nathan, Cody Jon, McCaw Morgan, Zhao Changzeng, Birchler James A
Division of Biological Sciences, University of Missouri, Columbia, MO, 65211, USA.
Methods Mol Biol. 2016;1469:1-13. doi: 10.1007/978-1-4939-4931-1_1.
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.
人工微型染色体是能够在主染色体组外稳定维持转基因的非整合载体。微型染色体的产生依赖于端粒介导的染色体截断，这涉及将转基因和端粒序列同时引入细胞以截断内源性染色体靶点。可采用两种方法；要么将端粒序列整合到二元载体中，用于根癌农杆菌转化，要么在粒子轰击过程中将端粒序列与转基因共同引入。在本方案中，介绍了分离和引入端粒序列所需的方法。按照所介绍的方法，可遵循标准转化程序来培育含有微型染色体的植物。
