Replication of mini-Mu prophage DNA.
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Mutations in rpsB that reduce the levels of the ribosomal protein S2 enhance the translation of cl in lambda lysogens. Two features of the cl transcript are required for enhanced translation: the absence of a leader and the presence of a downstream box, a sequence within the cl coding region that is complementary to the 16S rRNA. 30S ribosomal subunits deficient in S2 form ternary complexes with the cl transcript more efficiently than wild-type subunits. The absence of S2 may change the structure of the 16S rRNA, improving contacts with the cl downstream box.
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Electron microscopic analysis of heteroduplex molecules between the 94-kb plasmid p15B and the 92-kb phage P1 genome revealed nine regions of nonhomology, eight substitutions, and two neighboring insertions. Overall, the homologous segments correspond to 83% of the P1 genome and 81% of p15B. Heteroduplex molecules between p15B and the 99-kb phage P7 genome showed nonhomology in eight of the same nine regions; in addition, two new nonhomologous segments are present and P7 carries a 5-kb insertion representing Tn902. The DNA homology between those two genomes amounts to 79% of P7 DNA and 83% of p15B. Plasmid p15B contains two stem-loop structures. One of them has no equivalent structure on P1 and P7 DNA. The other substitutes the invertible C segments of P1 and P7 and their flanking sequences including cin, the gene for the site-specific recombinase mediating inversion.
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The inactivation kinetics of the lambda repressor following bleomycin (BM), UV-irradiation and nalidixic acid (NAL) treatments were studied in the recB21 mutant of E. coli K12. The results showed essentially normal induction by UV-irradiation, delayed induction by BM and no induction by NAL. The results were compared with inactivation kinetics in lexA1 and recF143 mutants.
Inactivation and mutagenesis of Escherichia coli by ultraviolet light and N-methyl-N-nitrosourea (MNU) were examined in isogenic strains that were (i) free of lambda phage, (ii) lysogenic for lambda or (iii) contained the lambda rexAB operon on a plasmid (prex20). Inactivation was enhanced when rex genes were present and this enhancement was greater for MNU than for UV. Mutagenesis was not greatly affected by the presence of the rex genes with one exception; UV-induced glnU degrees suppressor mutations were reduced in the strain harboring prex20. Control experiments using a strain containing only plasmid pACYC184 showed no effect on inactivation. A small depression in UV-induced mutagenesis was observed, however. These data suggest that the lambda RexAB proteins interfere with processes necessary for recovery from DNA damage and for fixation of one example of UV-induced mutation. This and other recent observations of effects by rex genes are discussed.
The Bacillus subtilis degSU two-gene operon encodes a two-component regulatory system which positively controls expression of a variety of genes encoding degradative enzymes. Point superactivator mutations in either degS or degU increase the production of these enzymes. A specialized transducing SP beta phage which partially complements several of these mutations was isolated from a phage library of the B. subtilis chromosome. This phage was shown to contain the degU (wild-type) gene. This unexpected co-dominance is discussed.
Infections of bacterial cultures by bacteriophages are common and serious problems in many biotechnological laboratories and factories. A method for specific, quantitative, and quick detection of phage contamination, based on the use of electric DNA chip is described here. Different phages of Escherichia coli and Bacillus subtilis were analyzed. Phage DNA was isolated from bacterial culture samples and detected by combination of bead-based sandwich hybridization with enzyme-labeled probes and detection of the enzymatic product using silicon chips. The assay resulted in specific signals from all four tested phages without significant background. Although high sensitivity was achieved in 4h assay time, a useful level of sensitivity (10(7)-10(8) phages) is achievable within 25 min. A multiplex DNA chip technique involving a mixture of probes allows for detection of various types of phages in one sample. These analyses confirmed the specificity of the assay.
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