通过脉冲场凝胶电泳对大肠杆菌染色体Tn5和F插入片段进行定位
Mapping of Escherichia coli chromosomal Tn5 and F insertions by pulsed field gel electrophoresis.
作者信息
Smith C L, Kolodner R D
机构信息
Department of Microbiology, College of Physicians and Surgeons, Columbia University, New York, New York 10032.
出版信息
Genetics. 1988 Jun;119(2):227-36. doi: 10.1093/genetics/119.2.227.
Abstract
A low resolution Not I physical map of Escherichia coli was recently constructed. In this report we demonstrated that this map can be used to map Tn5 and F insertions physically. The transposon, Tn5, contains Not I recognition sequences in its IS50 sequences. F plasmid contains an unmapped Not I site. Hence, the location of Tn5 and F in the chromosome can be mapped by identifying the location of the introduced Not I sites using pulsed field gel electrophoresis. The physical mapping of genetically mapped Tn5 insertions confirm the previously constructed Not I map and helps align the E. coli physical and genetic maps. The use of Tn5 can assist the construction of both physical and genetic maps for microorganisms lacking such maps. Variations on this approach will facilitate physical mapping with a wide variety of organisms, enzymes, and genetic elements.
摘要
最近构建了大肠杆菌的低分辨率Not I物理图谱。在本报告中，我们证明了该图谱可用于对Tn5和F插入片段进行物理定位。转座子Tn5在其IS50序列中含有Not I识别序列。F质粒含有一个未定位的Not I位点。因此，通过使用脉冲场凝胶电泳确定引入的Not I位点的位置，可对Tn5和F在染色体中的位置进行定位。对遗传定位的Tn5插入片段进行物理定位，证实了先前构建的Not I图谱，并有助于大肠杆菌物理图谱和遗传图谱的比对。使用Tn5有助于为缺乏此类图谱的微生物构建物理图谱和遗传图谱。这种方法的变体将促进对多种生物体、酶和遗传元件的物理定位。
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