Schmitt J M, Bohnert H J, Gordon K H, Herrmann R, Bernardi G, Crouse E J
Eur J Biochem. 1981 Jul;117(2):375-82. doi: 10.1111/j.1432-1033.1981.tb06348.x.
The chloroplast genomes of Euglena gracilis and Spinacia oleracea were investigated in their compositional heterogeneity, by using different experimental approaches. Euglena chloroplast DNA has a dG + dC content of 28%. Preparations averaging 20 x 10(6) in molecular weight exhibit a gross heterogeneity in their elution profiles from hydroxyapatite and in their buoyant densities because the rRNA genes have a high rG + rC content. Finer analysis by melting, buoyant density of restriction fragments and micrococcal nuclease degradation have revealed an extended compositional heterogeneity. From micrococcal nuclease digestion data, approximately 30% of the chloroplast genome is as low as 12% in its dG + dC content, whereas 10% is higher than 60% dG + dC. Since the average dG + dC content of large restriction endonuclease fragments varied to a lesser extent, most of dA + dT-rich sequences must occur in short stretches interspersed with dG + dC-rich stretches. Spinach chloroplast DNA (dG + dC = 36.5%) did not exhibit any gross compositional heterogeneity in its hydroxyapatite elution or in its buoyant density profile. But the higher resolution methods of melting, bouyant densities of restriction fragments and micrococcal nuclease degradation revealed a high degree of heterogeneity which appears to be due to interspersion of short DNA stretches of different base composition. About 30% of genome is as low as 22% in dG + dC, while 10% is higher than 60% in dG + dC.
利用不同的实验方法,对纤细裸藻和菠菜的叶绿体基因组组成异质性进行了研究。纤细裸藻叶绿体DNA的dG + dC含量为28%。平均分子量为20×10⁶的制剂在羟基磷灰石上的洗脱图谱和浮力密度方面表现出总体异质性,因为rRNA基因具有较高的rG + rC含量。通过解链、限制片段浮力密度和微球菌核酸酶降解进行的更精细分析揭示了广泛的组成异质性。根据微球菌核酸酶消化数据,约30%的叶绿体基因组dG + dC含量低至12%,而10%高于60% dG + dC。由于大的限制性内切酶片段的平均dG + dC含量变化较小,大多数富含dA + dT的序列必须以短片段形式散布于富含dG + dC的片段之间。菠菜叶绿体DNA(dG + dC = 36.5%)在羟基磷灰石洗脱或浮力密度图谱中未表现出任何总体组成异质性。但是,解链、限制片段浮力密度和微球菌核酸酶降解等更高分辨率的方法揭示了高度的异质性,这似乎是由于不同碱基组成的短DNA片段相互散布所致。约30%的基因组dG + dC含量低至22%,而10%高于60% dG + dC。