Ersland D R, Cattolico R A
Biochemistry. 1981 Nov 24;20(24):6886-93. doi: 10.1021/bi00527a023.
Nuclear DNA of the marine chromophytic alga Olisthodiscus luteus was analyzed in this study. Reassociation kinetics analysis has shown that 440-nucleotide DNA fragments from the genome of this alga contain 4% foldback, 58% repetitive, and 34% single-copy sequences. Precise analysis using isolated single-copy DNA revealed that Olisthodiscus has a large haploid DNA content of 1.66 x 10(-12) g/cell. For determination of the organization of single-copy and repetitive sequences within this genome, DNA fragments 3000 nucleotides in length were reassociated to C0t= 100 M . s. At this low C0t value 89% of the DNA bound to hydroxylapatite, suggesting that single-copy and repetitive elements are interspersed. The lengths of the duplexed repetitive DNA on these 3000-nucleotide fragments were measured by electron microscopy after digestion with S1 nuclease which removed the unreassociated single-copy DNA regions. Of these repetitive sequences, 68% were shorter than 1200 nucleotide pairs in length and had a modal length of 350 nucleotide pairs. A minor class of longer (to 4000 nucleotide pairs) repetitive sequences was also observed.
本研究分析了海洋色生性藻类黄褐异囊藻的核DNA。重缔合动力学分析表明,该藻类基因组中440个核苷酸的DNA片段包含4%的回文序列、58%的重复序列和34%的单拷贝序列。使用分离的单拷贝DNA进行的精确分析表明,黄褐异囊藻单倍体DNA含量很高,为1.66×10⁻¹²克/细胞。为了确定该基因组中单拷贝和重复序列的组织方式,将长度为3000个核苷酸的DNA片段重缔合至C0t = 100M·s。在这个低C0t值下,89%的DNA与羟基磷灰石结合,这表明单拷贝和重复元件是穿插分布的。在用S1核酸酶消化去除未重缔合的单拷贝DNA区域后,通过电子显微镜测量这些3000个核苷酸片段上双链重复DNA的长度。在这些重复序列中,68%的长度短于1200个核苷酸对,模式长度为350个核苷酸对。还观察到一小类较长(长达4000个核苷酸对)的重复序列。