Parker L L, Hall B G
Department of Molecular and Cell Biology, University of Connecticut, Storrs 06268.
Genetics. 1990 Mar;124(3):473-82. doi: 10.1093/genetics/124.3.473.
The cel (cellobiose utilization) operon of Escherichia coli K12 is not expressed in the wild-type organism. However, mutants that can express the operon and thereby utilize the beta-glucoside sugars cellobiose, arbutin and salicin are easily isolated. Two kinds of mutations are capable of activating the operon. The first involves mutations that allow the repressor to recognize the substrates cellobiose, arbutin and salicin as inducers. We have identified the sequence changes in five different active alleles and found those differences to be single base pair changes at one of two lysine codons in the repressor gene. The second kind of mutation involves the integration of the insertion sequences IS1, IS2 or IS5 into a 108-bp region 72-180 bp upstream of the start of transcription. Integration occurs at several different sites and in different orientations. Transcription of the cel operon begins at the same base pair in all mutants examined. Of 44 independent cel+ mutants, 27 were activated by point mutations and 17 were activated by insertion sequences. The preferred mechanism of activation appears to be strain dependent, since one of the parents yielded 94% insertionally activated alleles, while another yielded 100% point mutation activated alleles.
大肠杆菌K12的cel(纤维二糖利用)操纵子在野生型菌株中不表达。然而,能够表达该操纵子从而利用β-葡萄糖苷糖纤维二糖、熊果苷和水杨苷的突变体很容易分离得到。有两种突变能够激活该操纵子。第一种涉及使阻遏物将底物纤维二糖、熊果苷和水杨苷识别为诱导物的突变。我们已经确定了五个不同活性等位基因中的序列变化,发现这些差异是阻遏物基因中两个赖氨酸密码子之一处的单碱基对变化。第二种突变涉及插入序列IS1、IS2或IS5整合到转录起始点上游72 - 180 bp的一个108 bp区域。整合发生在几个不同的位点且方向不同。在所有检测的突变体中,cel操纵子的转录都从相同的碱基对开始。在44个独立的cel+突变体中,27个是由点突变激活的,17个是由插入序列激活的。激活的首选机制似乎取决于菌株,因为其中一个亲本产生了94%由插入激活的等位基因,而另一个亲本产生了100%由点突变激活的等位基因。