大肠杆菌K-12菌株ilvEDAC基因的物理组织。
Physical organization of the ilvEDAC genes of Escherichia coli strain K-12.
作者信息
McCorkle G M, Leathers T D, Umbarger H E
出版信息
Proc Natl Acad Sci U S A. 1978 Jan;75(1):89-93. doi: 10.1073/pnas.75.1.89.
Abstract
We have determined the physical location of the ilvEDAC genes on the restriction cleavage map of the ilv region of Escherichia coli K-12 by two methods: (i) heteroduplex and endonuclease cleavage analysis of hybrid phages carrying genetically defined parts of the ilv cluster and (ii) complementation analysis and enzyme assays to determine ilv gene expression from hybrid plasmids containing DNA restriction fragments of the transducing phage lambdah80dilv. The ilvEDA and ilvC operons occupy 2.4 and 0.9 megadalton sequences of DNA, respectively, and are separated by a region of 0.6-0.75 megadalton. The ilvD region, specifying dihydroxy acid dehydrase, has a maximum coding capacity of about 55,000 daltons of polypeptide. Our results confirm that ilvC is transcribed clockwise on the E. coli K-12 map, in the same direction as ilaEDA. A secondary lambda attachment site within ilvC has been located on a small (0.45 megadalton) EcoRI fragment. Our results are compared to other physical studies of ilv DNA.
摘要
我们通过两种方法确定了大肠杆菌K-12 ilv区域限制性酶切图谱上ilvEDAC基因的物理位置：（i）对携带ilv簇遗传定义部分的杂交噬菌体进行异源双链和核酸内切酶切割分析，以及（ii）进行互补分析和酶活性测定，以确定来自含有转导噬菌体λh80dilv DNA限制性片段的杂交质粒的ilv基因表达。ilvEDA和ilvC操纵子分别占据2.4和0.9兆道尔顿的DNA序列，中间相隔0.6 - 0.75兆道尔顿的区域。指定二羟酸脱水酶的ilvD区域，其多肽的最大编码能力约为55,000道尔顿。我们的结果证实，ilvC在大肠杆菌K-12图谱上是顺时针转录的，与ilaEDA方向相同。ilvC内的一个次要λ附着位点位于一个小的（0.45兆道尔顿）EcoRI片段上。我们将结果与其他关于ilv DNA的物理研究进行了比较。
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