Sulimova G E, Dokhiem M, Vaniushin B F
Mol Biol (Mosk). 1978 Jul-Aug;12(4):845-52.
The GC and AT fractions (GC content is 55.2 and 36.2 mole %, respectively) were isolated from wheat germ DNA by the thermal fractionation method (sequential thermal DNA denaturation followed by separation of denatured and undenatured DNA molecules by hydroxyapatitie chromatography). The amount of each of the two fractions isolated corresponds to about 25% of the total DNA. The 5-methylcytosine (m5C) content in the GC fraction (9.2 mole %) is 4.6 times higher than in the AT fraction. The DNA fractions isolated are very different in the level of methylation of cytosine residues (m5C/C+m5C ratio differs 3 times), in the amount of respective pyrimidine isostichs and in the renaturation kinetics. In contrast to the AT fraction, in the GC fraction the amount of dipyrimidine isostichs is higher than monopyrimidine ones. The AT and GC fractions isolated contain unique, moderately and highly reiterated nucleotide sequences. About 90% of the GC fraction is represented by reiterated sequences. The isolated (from the GC fraction) highly reiterated sequences (GC fraction I, Cot = 2.8 X 10(-2)) are very similar to the original GC fraction in the GC and m5C contents as well as in the amount of m5C in the respective pyrimidine isostichs. GC fraction I differs from the total GC fraction in a lower pyrimidine clustering degree. In total DNA and in GC and AT fractions m5C is found in mono- (34-40% of total m5C), di-(26-30%) and in all longer pyrimidine isostichs. It is suggested that in plants there exist many types of DNA sequences to be methylated as well as many respective DNA methylases.
通过热分级分离法(依次进行热DNA变性,然后通过羟基磷灰石色谱法分离变性和未变性的DNA分子)从小麦胚芽DNA中分离出GC和AT组分(GC含量分别为55.2和36.2摩尔%)。分离出的两种组分的量各自约占总DNA的25%。GC组分中的5-甲基胞嘧啶(m5C)含量(9.2摩尔%)比AT组分高4.6倍。分离出的DNA组分在胞嘧啶残基的甲基化水平(m5C/C + m5C比率相差3倍)、各自嘧啶同聚物的量以及复性动力学方面有很大差异。与AT组分相反,GC组分中双嘧啶同聚物的量高于单嘧啶同聚物。分离出的AT和GC组分包含独特的、中度和高度重复的核苷酸序列。GC组分中约90%由重复序列组成。从GC组分中分离出的高度重复序列(GC组分I,Cot = 2.8×10(-2))在GC和m5C含量以及各自嘧啶同聚物中的m5C量方面与原始GC组分非常相似。GC组分I与总GC组分的不同之处在于嘧啶聚类程度较低。在总DNA以及GC和AT组分中,m5C存在于单嘧啶(占总m5C的34 - 40%)、双嘧啶(26 - 30%)以及所有更长的嘧啶同聚物中。有人提出,在植物中存在许多类型的待甲基化DNA序列以及许多相应的DNA甲基化酶。