K部分：原核细胞转录.pptVIP

The polymerases of bacteriophage T3 and T7 are smaller single polypeptide chains, they synthesize RNA rapidly (200 nt/sec) and recognize their own promoters which are different from E. coli promoters. E. coli RNA polymerase E. coli polymerase: a subunit Two identical subunits in the core enzyme Encoded by the rpoA gene Required for assembly of the core enzyme Plays a role in promoter recognition Plays a role in the interaction of RNA polymerase with some regulatory factors b is encoded by rpoB gene, and b’ is encoded by rpoC gene . Make up the catalytic center of the RNA polymerase. Their sequences are related to those of the largest subunits of eukaryotic RNA polymerases. The b subunit can be cross linked to the template DNA, the product RNA, and the substrate ribonucleotides; mutations in rpoB affect all stages of transcription. Mutations in rpoC show that b’ also is involved at all stages. b’ subunit Binds two Zn 2+ ions and may participate in the catalytic function of the polymerase Hyparin (肝素)：binds to the b’ subunit and inhibits transcription in vitro. Hyparin competes with DNA for binding to the polymerase. 2. b’ subunit may be responsible for binding to the template DNA . Many prokaryotes contain multiple s factors to recognize different promoters. The most common s factor in E. coli is s70. Binding of the s factor converts the core RNA pol into the holoenzyme（全酶）. s factor is critical in promoter recognition（启动子识别）, by decreasing the affinity of the core enzyme for non-specific DNA sites (104) and increasing the affinity for the corresponding promoter. s factor is released from the RNA pol after initiation (RNA chain is 8-9 nt) Less amount of s factor is required in cells than that of the other subunits of the RNA pol. Consists of a sequence of between 40 and 60 bp -55 to +20: bound by the polymerase -20 to +20: tightly associated with the polymerase and protected from nuclease digestion by DNaseΙ. Up to position –40: critical for promoter functi