Vidal-Ingigliardi D, Raibaud O
Nucleic Acids Res. 1985 Feb 25;13(4):1163-72. doi: 10.1093/nar/13.4.1163.
Using in vitro techniques we have fused upstream sequences from the malPp promoter (normally activated by the MalT protein) to downstream sequences from the lacZp promoter (normally repressed by the LacI protein). Several hybrid promoters were thus obtained, which were controlled by the MalT protein, but were poorly active. More efficient promoters were then isolated using in vivo selection. Three main conclusions could be derived from the analysis of all of these hybrid promoters. Firstly, the MalT protein seems able to force RNA polymerase to start transcription at any DNA sequence, albeit with a low efficiency. Secondly, the strength of the hybrid promoters is considerably increased if a Pribnow Box is positioned at a precise location with respect to the MalT binding site. Thirdly, the presence of the lac operator, even when properly positioned with respect to the transcription startpoint, does not suffice to permit full repression by the lacI product.
我们使用体外技术将来自malPp启动子(通常由MalT蛋白激活)的上游序列与来自lacZp启动子(通常由LacI蛋白抑制)的下游序列融合。由此获得了几个杂合启动子,它们受MalT蛋白控制,但活性较低。然后利用体内筛选分离出了更有效的启动子。对所有这些杂合启动子的分析可得出三个主要结论。首先,MalT蛋白似乎能够迫使RNA聚合酶在任何DNA序列处起始转录,尽管效率较低。其次,如果普里布诺框(Pribnow Box)相对于MalT结合位点位于精确位置,则杂合启动子的强度会显著增加。第三,即使lac操纵子相对于转录起始点位置合适,其存在也不足以使lacI产物实现完全抑制。