Meister Armin
Department of Cytogenetics, Institute of Plant Genetics and Crop Plant Research, Corrensstr. 3, D-06466 Gatersleben, Germany.
J Theor Biol. 2005 Jan 7;232(1):93-7. doi: 10.1016/j.jtbi.2004.07.022.
From biochemical experiments it has been found that AT- and GC-specific dyes need a certain number of consecutive bases of the same type for binding one dye molecule. From known base sequences the amount of bases included in dye binding can be calculated and compared with experimental data from flow cytometry. Oryza sativa and Arabidopsis thaliana are the first higher plants which are nearly completely (>90%) sequenced. From the published sequences the theoretical fluorescence intensity of base-specific dyes in relation to a base-unspecific dye is calculated for different binding lengths. These values are compared with the actual fluorescence intensities of nuclei analyzed by flow cytometry. For all investigated dyes (DAPI, Hoechst 33258, Hoechst 33342 (all AT specific) and Mithramycin A (GC specific)) a binding length of 1 bp results from the comparison of theoretical and experimental data. This is, however, in disagreement with former results on dye binding. The main reason for the discrepancy seems to be the remaining gap in the sequencing of the Arabidopsis genome.
通过生化实验发现，AT特异性和GC特异性染料需要一定数量的相同类型的连续碱基才能结合一个染料分子。根据已知的碱基序列，可以计算出染料结合中包含的碱基数，并与流式细胞术的实验数据进行比较。水稻和拟南芥是首批几乎完全（>90%）测序的高等植物。根据已发表的序列，计算出不同结合长度下碱基特异性染料与碱基非特异性染料相关的理论荧光强度。将这些值与通过流式细胞术分析的细胞核的实际荧光强度进行比较。对于所有研究的染料（DAPI、Hoechst 33258、Hoechst 33342（均为AT特异性）和光神霉素A（GC特异性）），理论和实验数据的比较得出结合长度为1个碱基对。然而，这与以前关于染料结合的结果不一致。差异的主要原因似乎是拟南芥基因组测序中仍存在的缺口。
