Study of Pure Proteins, Nucleic Acids and Their Complexes from Halobacteria of the Dead Sea: RNA (Ribonucleic Acid) Polymerase-DNA Interaction

摘要

In the first year of this project we have isolated and cloned two rRNA operons of Halobacterium marismortui, each coding for 5s, 16s, and 23s rRNA genes. The structures of the two operons were compared by restriction mapping, by hybridization of different restriction fragments to phosphorus 32 labelled rRNA molecules and by R-loop analysis. The two operons are identical. The direction of transcription was determined to be 5'-16s-23s-5s-3' and tRNA genes were also identified. The promoter regions of the two operons as well as their 16s rRNA maturation sites were determined using S1 analysis. One operon has one promoter 270 bp upstream from the matured 16s rRNA and the other operon has three promoters in tandem at distances of 295-470bp upstream from the matured 16s rRNA. Sequencing of analysis at high salt concentration, for the characterization of the polymerase binding site in the promoter region. A major aim is to reconstitute a promoter dependent transcriptional assay.