Construction of rice mini-chromosomes by telomere-mediated chromosomal truncation.
Construction of rice mini-chromosomes by telomere-mediated chromosomal truncation.
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DOI：
10.1111/j.1365-313x.2012.04916.x
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发表时间：
2012-06
期刊：
影响因子：
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通讯作者：
Chunhui Xu;Zhukuan Cheng;Weichang Yu
中科院分区：
文献类型：
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作者：
Chunhui Xu;Zhukuan Cheng;Weichang Yu
端粒截短已被证明是一种有效的技术，用于创建微型染色体，可用作人工染色体平台的基因工程。基于人工染色体的基因工程被认为是上级于现有的通过农杆菌的随机基因整合或生物弹介导的遗传转化的技术。它将多个转基因组织成一个独特的遗传连锁块，用于随后的育种操作。端粒截短技术依赖于三个组成部分：介导染色体截短的端粒序列，允许选择转基因事件的选择标记，以及可用于通过基因靶向将未来基因接受到微型染色体中的位点特异性重组系统。这些元件通常在转换之前预先组装，这是一个既耗时又耗力的过程。本研究发现，这三种元素可以在基因枪转化中混合转化植物细胞，并在水稻中产生有效的染色体截短和微型染色体。该系统将允许快速构建具有抗性标记、位点特异性重组系统和其他期望元件的灵活选择的微型染色体。此外，使用水稻端三体系作为染色体截短的起始材料。从端部着丝粒染色体和其他染色体的截短中回收了微型染色体。在2年的继代培养中，微型染色体保持稳定。水稻是一种重要的经济作物，其微型染色体的构建将为未来基于人工染色体的水稻多基因叠加遗传工程提供平台。
Telomere truncation has been shown to be an efficient technology for the creation of mini-chromosomes that can be used as artificial chromosome platforms for genetic engineering. Artificial chromosome-based genetic engineering is considered to be superior to the existing techniques of randomized gene integration by Agrobacterium or biolistic-mediated genetic transformation. It organizes multiple transgenes as a unique genetic linkage block for subsequent manipulations in breeding. Telomere truncation technology relies on three components: the telomere sequence that mediates chromosomal truncation, a selection marker that allows the selection of transgenic events, and a site-specific recombination system that can be used to accept future genes into the mini-chromosome by gene targeting. These elements are usually pre-assembled before transformation, a process that is both time and labor consuming. We found in this research that the three elements could be mixed to transform plant cells in a biolistic transformation, and produced efficient chromosomal truncations and mini-chromosomes in rice. This system will allow rapid construction of mini-chromosomes with a flexible selection of resistant markers, site-specific recombination systems and other desirable elements. In addition, a rice telotrisomic line was used as the starting material for chromosomal truncations. Mini-chromosomes from the truncations of both the telocentric chromosome and other chromosomes were recovered. The mini-chromosomes remained stable during 2 years of subculture. The construction of mini-chromosomes in rice, an economically important crop, will provide a platform for future artificial chromosome-based genetic engineering of rice for stacking multiple genes.