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The structural changes in the cell membrane of λ-lysogenic Escherichia coli induced by colicin E2 were examined. The addition of colicin E2 made the cells susceptible to various detergents and the transport rate of o-nitrophenyl-β-D-galactoside into the colicin-treated cells was stimulated markedly by adding a low concentration of sodium dodecyl sulfate. The fluorescence intensity of 8-anilino-1-naphthalenesulfonate bound to the cells was markedly increased by adding colicin E2. Colicin E2 stimulated the incorporation of 32P from prelabeled phosphatidylglycerol to cardiolipin. All these changes probably suggesting the structural alteration of the cell membrane were dependent on the presence of the rex gene of λ prophage in the cells.  相似文献   

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Release of colicin E2 from Escherichia coli.   总被引:4,自引:3,他引:1       下载免费PDF全文
Treatment of Escherichia coli K-12(ColE2.P9) with 500 ng of mitomycin C per ml resulted in rapid and almost synchronous colicin E2 production. Colicin accumulated outside the cytoplasmic membrane, most probably in the periplasmic space. Colicin release occurred during a period in which the turbidity of the culture declined markedly. Periplasmic alkaline phosphatase was released during the same period, but cytoplasmic beta-galactosidase release was delayed.  相似文献   

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Plasmids were isolated from E colicinogenic strains and transformed into prototrophic Escherichia coli K 12 strain DB364. Screening of E colicinogenic transformants for growth on defined medium revealed an apparent amino acid auxotrophy mediated by E4 and, to a lesser extent, E7 colicin plasmids. The auxotrophy was further investigated in E4 colicinogenic strains. From such auxotrophic transformants, denoted Pmi+ (plasmid-mediated inhibition of growth), Pmi- variants were obtained at a frequency of 3 X 10(-4) per bacterium. Plasmid loss was not detected among Pmi- clones. Isolation of E4 colicin plasmids from Pmi- clones and retransformation of strain DB364 with these plasmids showed that 40% of the plasmids were unable to inhibit growth of DB364 and were inferred to have alterations in an E4 colicin plasmid gene termed pmi. All such plasmids were indistinguishable from native E4 colicin plasmids, with respect to colicin immunity, colicin production and excretion, and sensitivity to lysis by mitomycin C. Experiments examining the nutritional basis of the plasmid-mediated auxotrophy indicated that at least seven amino acids, isoleucine, leucine, valine, arginine, methionine, serine and glycine, were involved in the auxotrophy. However, supplementation with only these seven amino acids did not completely restore growth. Assays of the activities of enzymes involved in amino acid biosynthesis in colicinogenic and non-colicinogenic strains under repressing and derepressing growth conditions suggested that E4 colicin plasmids did not repress synthesis of the implicated amino acids.  相似文献   

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A new class of colicin sensitivity mutants of Escherichia coli was isolated whose cell division was specifically inhibited by colicin E(2) without detectable degradation of deoxyribonucleic acid (DNA) at 30 C. The mutant could not form colonies in the presence of colicin E(2) but recovered colony-forming ability by trypsin treatment even after prolonged incubation with the colicin. Addition of colicin E(2) to the exponentially growing mutant inhibited cell division completely but did not induce degradation of DNA into cold acid-soluble materials nor any breakage of DNA strands. Synthesis of DNA in the mutant was not inhibited, and long filamentous cells with multiple nuclear bodies were formed by the action of colicin E(2). Degradation of ribosomal ribonucleic acid and development of prophage lambda, both of which were induced by colicin E(2) in the sensitive cells, did not occur in the mutant. At the elevated temperature, however, the mutant was found to undergo colicin-induced degradation of DNA. No differences in ultraviolet light nor drug sensitivities were observed in the mutant compared to the parent E. coli. The data suggested that colicin E(2) had a specific inhibitory effect on cell division of E. coli that was not a consequence of DNA degradation.  相似文献   

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Fixation of colicin E3 to sensitive bacteria is followed, after a lag of 2 to 6 min, by the rapid degradation of all the RNA of the 30S ribosomal subunits, yielding a large 15.5S fragment and a smaller fragment, containing the 3′-terminal end of the 16S RNA. The small RNA fragment which was estimated to consist of about 52 nucleotides, was retained within the 30S subunit in vivo and was subsequently recovered quantitatively without apparent further degradation. Kinetic studies of the cleavage of 16S RNA indicated that this is the primary and lethal effect of colicin E3 and the primary cause of the observed inhibition of protein synthesis in vivo. Small amounts of an RNA fragment, apparently identical in size to the small E3-fragment, were also isolated from 30S particles obtained from untreated bacteria.  相似文献   

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Escherichia coli strain CL137, a K-12 derivative made E colicinogenic by contact with Fredericq's strain K317, was unaffected by colicin E2-P9, but K-12 carrying ColE2-P9 was sensitive to the E colicin made by strains CL137 and K317. This colicin we named E7-K317 because by the test of colicinogenic immunity it differed from colicins E1-K30, E2-P9, and E3-CA38 and from recently recognized colicins termed E4Horak, E5, and E6. Strain K317 as conjugational donor transmitted E7 colicinogeny; about half the E7-colicinogenic transconjugants were immune to colicin E2-P9. A spontaneous variant of CL137 retained E7 colicinogeny but was sensitive to E2 colicins. We attribute the E2 immunity of strain CL137 and some E7-coliconogeic transconjugants to a "colicin-immunity plasmid," ColE2imm-K317, from strain K317. Tra+ E7-colicinogenic transconjugants restricted phage BF23 in the same way as strains carrying ColIb-P9. We attribute Tra+ and restricting ability to a plasmid, pRES-K317, acquired from strain K317, and related to the ColI plasmids.  相似文献   

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