Di Laurenzio L, Frost L S, Paranchych W
Department of Biochemistry, University of Alberta, Edmonton, Canada.
Mol Microbiol. 1992 Oct;6(20):2951-9. doi: 10.1111/j.1365-2958.1992.tb01754.x.
The gene encoding the TraM protein of the conjugative plasmid F was cloned, overexpressed and the gene product was purified. The TraM protein was found in the cytoplasm of cells carrying the F plasmid with a smaller amount in the inner membrane. DNase I footprinting experiments showed that the purified protein protects three regions in the F oriT locus with different affinity for the upper and lower strands of DNA. A 15-nucleotide motif was identified within the protected regions that represented the DNA-binding site. The TraM protein was also found to bind to a sequence in the oriT region of the non-conjugative plasmid ColE1 that resembles the three binding sites in the F oriT region.
对接合质粒F的TraM蛋白编码基因进行了克隆、过表达,并对基因产物进行了纯化。在携带F质粒的细胞胞质中发现了TraM蛋白,内膜中的含量较少。DNA酶I足迹实验表明,纯化后的蛋白以不同亲和力保护F oriT基因座中的三个区域,分别针对DNA的上链和下链。在代表DNA结合位点的受保护区域内鉴定出一个15个核苷酸的基序。还发现TraM蛋白与非接合质粒ColE1的oriT区域中的一个序列结合,该序列类似于F oriT区域中的三个结合位点。