[Study of the binding of RNA polymerase by a recombinant plasmid using electron microscopy].
RNA-polymerase of E. coli was bound in vitro under physiological conditions to a recombinant plasmid pBR322 carrying two identical segments of bacteriophage T4 DNA, each containing a complete gene coding for T4 DNA ligase. After fixation of the complex with formaldehyde it was analysed by electron microscopy. A map of binding sites of the enzyme to DNA was obtained after a statistical assessment of micrographs. The inserted repeat revealed itself in the map as two regions of identical binding patterns, thus proving the adequacy of the preparation procedure. In pBR322 the strong binding sites correlate with the position of promoters. Also, apart from this, there are other strong binding sites within the T4 sequences which correlate strongly with the regions of abnormally high AT content. That means that under physiological conditions the RNA polymerase forms strong binding complexes with any AT rich DNA regions, as well as with real promoters.
Beritashvili
D R
DR
Kalandadze
A G
AG
Kraev
A S
AS
D001665:Binding Sites; D002853:Chromatography, Liquid; D004247:DNA / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology; D008854:Microscopy, Electron; D010957:Plasmids; D011485:Protein Binding; D011995:Recombination, Genetic; D013604:T-Phages / Q000235:genetics
D004247:DNA; D012321:DNA-Directed RNA Polymerases
Izuchenie sviazyvaniia rnk-polimerazy s rekombinantnoĭ metodom élektronnoĭ mikroskopii.