Teo Chee How, Lermontova Inna, Houben Andreas, Mette Michael Florian, Schubert Ingo
Leibniz Institute of Plant Genetics and Crop Plant Research IPK, Corrensstrasse 3, 06466 Gatersleben, Germany.
Chromosoma. 2013 Jun;122(3):233-41. doi: 10.1007/s00412-013-0406-0. Epub 2013 Mar 23.
Artificial minichromosomes are highly desirable tools for basic research, breeding, and biotechnology purposes. We present an option to generate plant artificial minichromosomes via de novo engineering of plant centromeres in Arabidopsis thaliana by targeting kinetochore proteins to tandem repeat arrays at non-centromeric positions. We employed the bacterial lactose repressor/lactose operator system to guide derivatives of the centromeric histone H3 variant cenH3 to LacO operator sequences. Tethering of cenH3 to non-centromeric loci led to de novo assembly of kinetochore proteins and to dicentric carrier chromosomes which potentially form anaphase bridges. This approach will be further developed and may contribute to generating minichromosomes from preselected genomic regions, potentially even in a diploid background.
人工微型染色体是用于基础研究、育种和生物技术目的的非常理想的工具。我们提出了一种通过在拟南芥中对植物着丝粒进行从头工程设计来生成植物人工微型染色体的方法，即将动粒蛋白靶向到非着丝粒位置的串联重复阵列上。我们利用细菌乳糖阻遏物/乳糖操纵子系统，将着丝粒组蛋白H3变体cenH3的衍生物引导至LacO操纵序列。将cenH3拴系到非着丝粒位点会导致动粒蛋白的从头组装，并产生可能形成后期桥的双着丝粒载体染色体。这种方法将得到进一步发展，可能有助于从预选的基因组区域生成微型染色体，甚至可能在二倍体背景下实现。
