Oliphant A R, Struhl K
Department of Biological Chemistry, Harvard Medical School, Boston, MA 02115.
Nucleic Acids Res. 1988 Aug 11;16(15):7673-83. doi: 10.1093/nar/16.15.7673.
The consensus sequence of E.coli promoter elements was determined by the method of random selection. A large collection of hybrid molecules was produced in which random-sequence oligonucleotides were cloned in place of a wild-type promoter element, and functional -10 and -35 E.coli promoter elements were obtained by a genetic selection involving the expression of a structural gene. The DNA sequences and relative levels of function for -10 and -35 elements were determined. The consensus sequences determined by this approach are very similar to those determined by comparing DNA sequences of naturally occurring E.coli promoters. However, no strong correlation is observed between similarity to the consensus and relative level of function. The results are considered in terms of E.coli promoter function and of the general applicability of the random selection method.
通过随机选择的方法确定了大肠杆菌启动子元件的共有序列。构建了大量的杂交分子,其中随机序列的寡核苷酸被克隆以取代野生型启动子元件,并通过涉及结构基因表达的遗传筛选获得了功能性的大肠杆菌-10和-35启动子元件。测定了-10和-35元件的DNA序列及相对功能水平。通过这种方法确定的共有序列与通过比较天然存在的大肠杆菌启动子的DNA序列所确定的序列非常相似。然而,在与共有序列的相似性和相对功能水平之间未观察到强相关性。从大肠杆菌启动子功能以及随机选择方法的普遍适用性方面对结果进行了考量。