Palmer J D
Nucleic Acids Res. 1982 Mar 11;10(5):1593-605. doi: 10.1093/nar/10.5.1593.
A rapid and simple method for constructing restriction maps of large DNAs (100-200 kb) is presented. The utility of this method is illustrated by mapping the Sal I, Sac I, and Hpa I sites of the 152 kb Atriplex triangularis chloroplast genome, and the Sal I and Pvu II sites of the 155 kb Cucumis sativa chloroplast genome. These two chloroplast DNAs are very similar in organization; both feature the near-universal chloroplast DNA inverted repeat sequence of 22-25 kb. The positions of four different genes have been localized on these chloroplast DNAs. In both genomes the 16S and 23S ribosomal RNAs are encoded by duplicate genes situated at one end of the inverted repeat, while genes for the large subunit of ribulose-1,5-bisphosphate carboxylase and a 32 kilodalton photosystem II polypeptide are separated by 55 kb of DNA within the large single copy region. The physical and genetic organization of these DNAs is compared to that of spinach chloroplast DNA.
本文介绍了一种快速简便的构建大DNA(100 - 200 kb)限制性图谱的方法。通过绘制152 kb的三角叶滨藜叶绿体基因组的Sal I、Sac I和Hpa I酶切位点,以及155 kb的黄瓜叶绿体基因组的Sal I和Pvu II酶切位点,说明了该方法的实用性。这两个叶绿体DNA在组织结构上非常相似;两者都具有22 - 25 kb的近乎通用的叶绿体DNA反向重复序列。四个不同基因的位置已定位在这些叶绿体DNA上。在这两个基因组中,16S和23S核糖体RNA由位于反向重复一端的重复基因编码,而1,5 - 二磷酸核酮糖羧化酶大亚基和32千道尔顿光系统II多肽的基因在大单拷贝区域内被55 kb的DNA隔开。将这些DNA的物理和遗传组织与菠菜叶绿体DNA的进行了比较。