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ϕX174 rolling circle replication Fig. 19.3 ϕX174 • + strand ssDNA genome, 5386 nt with 10 genes • Icosahedral capsid, attaches to LPS • Genome contains overlapping genes: some regions of polycistronic mRNA can be translated in >1 reading frame Many phages use this strategy to produce progeny genomes ϕX174 CisA nicks + strand of RF I dsDNA, remains attached to 5ʼ end; 3ʼOH of nicked strand primes synthesis (but no “lagging strand synthesis” for this phage) Fig. 19.4 Infection by T4 (T even phages) • • • • • Fig. 10.10 T4 binds to LPS in OM (recognition by tail fibers) 169 kd linear dsDNA is injected into cell from head by tail Capsid remains outside cell 200 phage in 25ʼ lytic cycle Genomes of T2, T4, and T6 are 85% identical, with differences accounting for binding different host receptors Program of T4 lytic cycle Fig. 10.15 Early phage proteins subvert host RNA synthesis (modifying host σ) and degrade host DNA, allowing phage to take over host cell; Specific phage DNA synthesis proteins. Late proteins: capsids assembled in 3 different paths. 1 Replication of T4 DNA, part I By 4 min post-infection, viral DNA pol initiates viral DNA replication. Viral DNA pol and recombination proteins produce complex intermediates--linear repeats called concatemers, that subsequently are processed into progeny genomes. Processing T4 genomic DNA • Replication & recombination yields long concatemers • Phage endonuclease processes DNA into linear molecules of specific length =105% of genome (no cutting at specific sequence) • Terminal redundancy of varying sequence called circular permutation Fig. 10.13 λ: lysis or lysogeny Phage λ • Linear dsDNA genome, 48.5Kb • Simpler tail structure than T even phages • Binds to maltoporin in OM of E. coli • Temperate phage, capable of 2 distinct fates upon infection Temperate phages can proceed with lytic infection or can integrate into genome and maintain prolonged association with host. Fig. 10.16 2 Forming a λ lysogen • λ-encoded Integrase catalyzes site-specific recombination with E. coli chromosome between gal-bio • Maintenance of lysogeny requires continued expression of cI repressor • Reversible process… Fig. 10.18 3