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  • Escherichia coli  (36)
  • Springer  (36)
  • American Chemical Society
  • Essen : Verl. Glückauf
  • Krefeld : Geologischer Dienst Nordhein-Westfalen
  • 2015-2019
  • 2005-2009
  • 1985-1989  (36)
  • 1980-1984
  • 2007
  • 1987  (36)
Collection
Publisher
Years
  • 2015-2019
  • 2005-2009
  • 1985-1989  (36)
  • 1980-1984
Year
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Journal of molecular evolution 24 (1987), S. 205-211 
    ISSN: 1432-1432
    Keywords: Endosymbiont ; Aphid ; Genome size ; Nucleotide composition ; Cell organelle ; Mycoplasma ; Escherichia coli
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary An intracellular symbiont was isolated from the mycetocyte of the pea aphidAcyrthosiphon pisum, and its genomic DNA was compared with those ofEscherichia coli andMycoplasma capricolum with respect to nucleotide composition and kinetic complexity. Thermal dissociation, CsCl density equilibrium centrifugation, and high-performance liquid chromatography of the nuclease P1 digest all indicated that the G+C content of the endosymbiont DNA is as low as 30%. In this respect, the endosymbiont resembledMycoplasma species. The reassociation kinetics of genomic DNA labeled by nick translation suggested that the endosymbiont genome is 1.4×1010 daltons in size, about 5 and 18 times as large as those ofE. coli andM. capricolum, respectively. The results were confirmed by reassociation of endosymbiont DNA labeled by incubation with [3Hthymidine in Grace's medium. The endosymbiont genome of the aphid was about 500 times larger than those of leafhopper endosymbionts previously analyzed by ultracentrifugation. These characteristic properties of the aphid endosymbiont genome are discussed in connection with the evolution of cell organelles, and with reference to a previous finding that most of the genes of the aphid endosymbiont are not expressed when present intracellularly.
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  • 2
    ISSN: 1432-072X
    Keywords: Creatinine deimination enzymes ; Creatinine deiminase ; Cytosine deaminase ; Creatinine degradation ; N-Methylhydantoin ; Creatinine ; Cytosine ; Pseudomonas putida 77 ; Cytosine deaminase induction ; Escherichia coli
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Creatinine deimination has been newly detected in the following various cytosine deaminase-forming microorganisms: Escherichia coli, Proteus mirabilis, Pseudomonas aureofaciens, Pseudomonas chlororaphis and Pseudomonas cruciviae. All these microorganisms, except for E. coli, formed cytosine deaminase in a constitutive or repressive way. P. putida 77 and E. coli showed highly increased formation of creatinine deiminase in the presence of creatinine and cytosine. Throughout serial DEAE-Sephacel and Sephacryl S-300 column chromatographies, the cytosine deaminases of these microorganisms, except for that of P. ovalis, were found to hydrolyze both creatinine and cytosine at comparable rates. No concrete evidence was obtained for the presence of any other protein that hydrolyzed creatine and/or cytosine than the cytosine deaminases in the three test microorganisms randomly selected for investigation. Different from P. putida 77, none of the test microorganisms degraded N-methylhydantoin; neither N-methylhydantoin amidohydrolase nor N-carbamoylsarcosine amidohydrolase was formed in the presence of creatinine in these microorganisms. As a result, the wide occurrence of cytosine deaminases in microorganisms was found to be related to the wide distribution of those microorganisms which hydrolyze creatinine to N-methylhydantoin without further degradation.
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  • 3
    ISSN: 1432-072X
    Keywords: F1F0 ATP synthase ; Escherichia coli ; Klebsiella pneumoniae ; Phylogenetic relationship
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The ATP synthase complex of Klebsiella pneumoniae (KF1F0) has been purified and characterized. SDS-gel electrophoresis of the purified F1F0 complexes revealed an identical subunit pattern for E. coli (EF1F0) and K. pneumoniae. Antibodies raised against EF1 complex and purified EF0 subunits recognized the corresponding polypeptides of EF1F0 and KF1F0 in immunoblot analysis. Protease digestion of the individual subunits generated an identical cleavage pattern for subunits α, β, γ, ε, a, and c of both enzymes. Only for subunit δ different cleavage products were obtained. The isolated subunit c of both organisms showed only a slight deviation in the amino acid composition. These data suggest that extensive homologies exist in primary and secondary structure of both ATP synthase complexes reflecting a close phylogenetic relationship between the two enterobacteric tribes.
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  • 4
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    Archives of microbiology 149 (1987), S. 36-42 
    ISSN: 1432-072X
    Keywords: Catabolite repression ; Genetics ; Malate dehydrogenase ; Molecular cloning ; Sequence ; CRP binding site ; Escherichia coli
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The malate dehydrogenase gene of Escherichia coli, which is susceptible to catabolite and anaerobic repression, has been cloned using plasmic pLC32-38 of Clarke and Carbon (1976). The nucleotide sequence was determined of a 2.47 kbp fragment, containing the mdh structural gene. All information necessary for expression of the mdh structural gene was mapped within a 1.3 kbp SphI-BstEII fragment. Compared with the untransformed wild type, transformations with pUC19 vector, containing this fragment, gave up to 40-fold more malate dehydrogenase activity in both E. coli wild type and mdh mutant recipients. Catabolite repression was not affected in the transformants. A possible CRP binding site in the promotor region of the mdh gene provides evidence for a co-regulation with fumA gene, the structural gene of fumarase, which is also subject to catabolite repression. The structures for transcription initiation and termination were similar to those previously described for E. coli. Amino acid sequence homologies between pro- and eucaryotic malate dehydrogenases are discussed.
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  • 5
    ISSN: 1432-072X
    Keywords: Escherichia coli ; Osmoregulation ; Glycine betaine ; Proline betaine ; γ-Butyrobetaine ; Trehalose ; Glutamic acid ; Nuclear magnetic resonance spectroscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract It has been shown previously that externally added glycine betaine is accumulated in Escherichia coli in response to the external osmotic strength. Here we have shown, by using nuclear magnetic resonance spectroscopy and radiochemical methods, that E. coli growing in a glucose-mineral medium of elevated osmotic strength generated with NaCl, had the same capacity to accumulate proline betaine and glycine betaine. Its capacity to accumulate γ-butyrobetaine was, however, 40 to 50% lower. Accordingly, externally added proline betaine and glycine betaine stimulated aerobic growth of osmotically stressed cells equally well, and they were more osmoprotective than γ-butyrobetaine. In cells grown at an osmotic strength of 0.64, 1.01, or 1.47 osmolal, respectively, the molal cytoplasmic concentration of the two former betaines corresponded to 29, 38, or 58% of the external osmotic strength. Nuclear magnetic resonance spectroscopy revealed that trehalose and glutamic acid were the only species of organic osmolytes accumulated in significant amounts in cells grown under osmotic stress in glucosemineral medium without betaines. Their combined molal concentration in the cytoplasm of cells grown at 1.01 osmolal corresponded to 27% of the external osmotic strength.
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  • 6
    ISSN: 1476-5535
    Keywords: Recombinant human insulin-like growth factor ; Escherichia coli ; Fermentation ; Production ; Somatomedin C
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Two kinds of fed batch fermentation processes were compared at a 10-liter scale to examine their effect on recombinant human insulin-like growth factor (IGF-1) gene expression inEscherichia coli. The difference between the two processes was the feed medium composition and whether the process used a single or dual feed during the course of the fermentation. In the dual feed system, organic nitrogen was delivered at a higher rate (50 g/h) than in the single feed system (5 g/h). The dual feed process resulted in a significant increase in IGF-1 yield. 30 mg IGF-1/g dry cell weight was synthesized in the dual feed system compared to 3 mg IGF-1/g dry cell weight obtained in the single feed system. The presence of high levels of organic nitrogen during the induction period may enhance IGF-1 synthesis by protecting the IGF-1 from proteolytic degradation. The IGF-1 yield decreased to 17 mg/g dry cell weight when the glucose supply was decreased from 17 g/h to 8 g/h during the induction period; however, an increase in glucose supply from 17 g/h to 50 g/h during the induction period did not enhance the IGF-1 synthesis. Thus, the enhancement of IGF-1 gene expression in the dual feed process was mainly dependent on a high level of organic nitrogen and an appropriate level of glucose in the medium during the induction period.
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  • 7
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    Molecular genetics and genomics 208 (1987), S. 485-489 
    ISSN: 1617-4623
    Keywords: PhoE porin ; Escherichia coli ; Insertion mutagenesis ; Topology ; Biogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary To study the effects of small insertions on the biogenesis and functioning of outer membrane pore protein PhoE of Escherichia coli K12, oligonucleotides were inserted at five different sites in the phoE gene. The proteins encoded by the mutant alleles all appeared to be incorporated into the outer membrane since cells producing these proteins bound either phages specific for the PhoE protein or monoclonal antibodies or both. However, one mutant protein was apparently not incorporated normally since expression of this protein was lethal and the protein did not function as a pore. Amino acid residues 75 and 280 of the PhoE protein appear to be exposed on the cell surface because insertions at these sites interfere with the binding of PhoE-specific phages or monoclonal antibodies to whole cells, respectively.
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  • 8
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    Molecular genetics and genomics 208 (1987), S. 511-517 
    ISSN: 1617-4623
    Keywords: Escherichia coli ; pH ; Outer membrane proteins ; Protein and operon fusions ; Osmosensor EnvZ
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Electrophoretic analysis of total membrane proteins of Escherichia coli cells grown at neutral or acid pH showed that ompF, ompC and lamB porin gene expression was regulated by changes in extracellular pH. Growth at acid pH was correlated with a decrease in outer membrane proteins OmpF and LamB and an increase in protein OmpC. pH-induced effects were confirmed and quantitatively estimated by using strains carrying ompF-lacZ, ompC-lacZ and lamB-lacZ fusions. Our studies showed that the pH-dependent regulation acted at a transcriptional level on ompF and ompC gene expression and also at a post-transcriptional level on ompF gene expression. Similar studies carried out with envZ strains showed that the pH-controlled transducing signal was mediated via the EnvZ protein, although other pH-dependent pathways were also involved.
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  • 9
    ISSN: 1617-4623
    Keywords: Escherichia coli ; RNA polymerase ; rpoH ; Heat-shock ; Multiple promoters
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Transcriptional start sites of the rpoH gene which codes for a minor σ factor (σ32) of Escherichia coli RNA polymerase were determined. The rpoH gene is transcribed, both in vivo and in vitro, from two major (P1 and P2) and one minor (P2*) promoters. In vitro synthesis of the rpoH mRNAs is dependent on the major species of RNA polymerase holoenzyme (Eσ70) but not on the minor one (Eσ32). S1 nuclease analysis of the in vivo RNA showed that the level of rpoH transcript from the downstream P2 promoter increases rapidly when E. coli cells are transferred from 30° C to 42° C, while the transcript from the upstream P1 promoter remains at a constant level. Under these conditions, the metabolic stabilities of rpoH mRNAs are virtually unaffected, suggesting that the synthesis of rpoH mRNA from the P2 promoter is specifically enhanced upon heatshock.
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  • 10
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    Molecular genetics and genomics 208 (1987), S. 288-293 
    ISSN: 1617-4623
    Keywords: Tellurium resistance ; Inducible ; Plasmid ; Escherichia coli
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A large (〉250 kb) conjugative plasmid, pMER610, specifying resistance to tellurium and mercury was isolated from an Alcaligenes strain and transferred by conjugation to Escherichia coli AB1157. The acquisition of pMER610 by AB1157 increased the resistance to both tellurite and tellurate by 100-fold. Expression of tellurite resistance by pMER610 and the cloned Ter determinant was inducible by prior exposure to tellurite at levels sub-toxic to the sesitive AB1157. Physical analysis of the cloned Ter fragment located the resistance determinant to a 3.55 kb region. Insertion of Tn 1000 (γδ) into this region produced two classes of sensitive mutations, fully sensitive and intermediate or hyposensitive, which map in adjacent regions and form two complementation groups. Maxicell analysis identified four polypeptides (15.5, 22, 23 and 41 kDa) expressed by the Ter clone. The 23 kDa polypeptide may not be required for resistance since tellurium-sensitive γδ insertion mutations were not detected in the 23 kDa coding region.
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  • 11
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    Molecular genetics and genomics 210 (1987), S. 504-508 
    ISSN: 1617-4623
    Keywords: Escherichia coli ; Cell division ; ftsE ; Operon ; Membrane location
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The cell division genes ftsY, ftsE and ftsX form an operon mapping at 76 min on the Escherichia coli chromosome. The protein products of these genes have been indentified previously. We have studied the cellular location of the radiolabelled Fts proteins using maxicells and standard fractionation procedures. Previous protein sequence homologies suggested an inner membrane location for FtsE. We have confirmed this predicted location and have shown that FtsY and FtsX are also inner membrane-associated. These results are igreement with the hypothesis that FtsE may act at the inner membrane, in a “septalsome” complex, by coupling ATP hydrolysis to the process of bacterial cell division.
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  • 12
    ISSN: 1617-4623
    Keywords: nuA operon ; Regulation ; RNA polymerase ; Escherichia coli
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The rate of synthesis and intracellular content of the NusA protein, a transcription termination factor, were determined for wild-type and nusA and/or nusB mutants of Escherichia coli. Both the rate and content of NusA in wild-type strains were similar to that of the RNA polymerase σ subunit, a transcription initiation factor, on a molar basis, and about 30%–40% the levels of RNA polymerase ββ′ subunits. At the stationary phase of cell growth, the values increased in parallel for both transcription factors up to approximately the level of the ββ′ subunits. In nus mutants, the rate of synthesis and the content of the σ subunit were significantly increased. These observations together suggest that the two transcription factors are coordinately regulated.
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  • 13
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    Molecular genetics and genomics 209 (1987), S. 188-192 
    ISSN: 1617-4623
    Keywords: oriC ; Escherichia coli ; Density shift ; Interreplication time ; Rate-limiting event
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The variability of the time interval between successive rounds of chromosome replication was estimated by density-shift experiments, by measuring the conversion of heavy DNA to hybrid density and light DNAs upon transfer of a steady-state culture growing in medium with [13C]glucose and 15NH4Cl to medium with light isotopes. The coefficient of variation (CV%) for the interreplication time of the Escherichia coli K12 chromosome was found to be 17%, i.e. similar to that for interdivision time. The presence of additional copies of oriC in the cell on a high copy number plasmid did not increase the CV of interreplication time. It is concluded that a single rate-limiting event is unlikely to time the initiation of chromosome replication. The regulation of initiation at oriC and the coordination with cell division is discussed.
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  • 14
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    Molecular genetics and genomics 209 (1987), S. 154-158 
    ISSN: 1617-4623
    Keywords: Recombination ; Plasmid ; Streptomyces ; Escherichia coli
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary An Escherichia coli-Streptomyces shuttle plasmid pIF132 containing two direct mel repeats was constructed. While pIF132 replicated relatively stably in E. coli (Rec+ or recA), its structure was unstable in S. lividans: recombination between the mel repeats resulted in a smaller plasmid, pIF138. Furthermore, pIF132 formed oligomers extensively in E. coli but not in S. lividans.
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  • 15
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    Molecular genetics and genomics 208 (1987), S. 70-75 
    ISSN: 1617-4623
    Keywords: Proline carrier gene ; putP ; Escherichia coli ; DNA sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The nucleotide sequence of the putP gene coding for the proline carrier in Escherichia coli has been determined and the amino acid sequence of the proline carrier deduced from it. The proline carrier is predicted to consist of 502 amino acids, resulting in a molecular weight of 54,343. The predicted protein is very hydrophobic (70% nonpolar amino acids), and its hydropathy profile suggests that it is composed of 12 hydrophobic segments with a mean length of 24.4 residues/segment. If these segments are assumed to be α-helical, the mean length of each domain corresponds to the thickness of the hydrophobic core of the membrane. Potential promoter, catabolite gene activator protein (CAP) binding sites and several palindromic sequence, which might be regulatory regions by the putA gene product, were also found in the 5′ flanking region of the postulated putP gene. A typical p-independent transcription termination signal was found after the terminator codon of the putP gene.
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  • 16
    ISSN: 1617-4623
    Keywords: Outer membrane permeability ; Outer membrane proteins ; TraT protein ; Escherichia coli ; Salmonella typhimurium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two insertion mutations, each introducing a new negatively charged amino acid residue into a hydrophobic region of the TraT protein coded by the F-like plasmid R6-5 caused significant alterations in outer membrane permeability of Escherichia coli and Salmonella typhimurium, so that strains carrying plasmids with either of these mutations became sensitive to hydrophobic antibiotics.
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  • 17
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    Molecular genetics and genomics 206 (1987), S. 356-357 
    ISSN: 1617-4623
    Keywords: tRNA 1 Asp ; aspV ; Escherichia coli ; dnaQ
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We report a new tRNA 1 Asp gene near the dnaQ gene, which is located at 5 min on the Escherichia coli linkage map. We named it aspV. The sequence corresponding to the mature tRNA is identical with that of the two previously identified tRNA 1 Asp genes (aspT and aspU), but there is no homology in the sequences of their 3′-and 5′-flanking regions.
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  • 18
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    Molecular genetics and genomics 206 (1987), S. 246-251 
    ISSN: 1617-4623
    Keywords: Iron transport ; Escherichia coli ; exbB gene ; Colicin uptake
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The exbB locus of Escherichia coli is involved in the uptake of certain iron(III) siderophore compounds, of vitamin B12 and of certain colicins. Outer membrane receptor proteins are essential constituents of the corresponding uptake systems. The DNA carrying the exbB locus was cloned into pACYC184 and subcloned into pUC18. With the use of insertion mutagenesis employing transposon Tn1000 and by deletion analysis, the exbB locus was confined to a 1.9 kb DNA fragment. An in vitro transcription/translation system and minicells programmed by exbB + plasmids expressed a protein with an apparent molecular weight of 26,000. One plasmid, designated pKE7, expressed this protein to an extent that it became a prominent band in the membrane fraction of transformants. In contrast, chromosomally encoded ExbB protein could not be detected. The plasmid-encoded ExbB protein was mainly localized in the cytoplasmic membrane. Ferrichrome transport in exbB mutants was restored by exbB + plasmids. Moderate overexpression of ExbB resulted in an enhanced ferrichrome transport, strong overexpression reduced the transport rate compared to a wild-type strain. The ExbB function shares some properties with the TonB function.
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  • 19
    ISSN: 1617-4623
    Keywords: N-methyl-N′-nitro-N-nitrosoguanidine ; Mutagenesis ; Lambda phage ; Escherichia coli
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The methylating agent N-methyl-N′-nitro-N-nitrosoguanidine preferentially induces G:C to A:T transitions at DNA base pairs with the G in one particular strand of the cI gene in a lambda prophage, in this case the non-transcribed straind, in Escherichia coli cells in which the adaptive response is induced. The same preference is found for the cI gene inserted in the genome in the inverse orientation, so the differential effect is not caused by the direction of motion of the DNA replicating fork.
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  • 20
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    Molecular genetics and genomics 209 (1987), S. 408-410 
    ISSN: 1617-4623
    Keywords: Escherichia coli ; HU-2 protein ; Gene cloning ; Nucleotide sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The Escherichia coli HU-2 gene was cloned using a DNA fragment from the HU-1 gene as a probe. The amino acid sequence of the HU-2 protein deduced from the nucleotide sequence is in good agreement with the published sequence. The nucleotide sequence has a possible promoter and a typical ribosomal binding site upstream of the translation initiation codon (AUG) and a possible rhoindependent terminater site downstream of the termination codon (UAA) of the gene.
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  • 21
    ISSN: 1617-4623
    Keywords: Escherichia coli ; RNA polymerase ; Sigma factor ; Heat-shock ; Synthetic peptide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Antibodies were raised against a synthetic tetradecameric peptide with an amino acid sequence, DLIQEGNIGLMKAV, which corresponds to the most highly conserved region of bacterial RNA polymerase σ factors. In a Western-blot analysis of total Escherichia coli proteins, the antiserum reacted specifically with at least three proteins with apparent molecular weights of 75 kDa, 27 kDa and 23 kDa, in addition to the known σ factors (σ70 and σ32). The majorities of σ70 and σ32 were recovered as associated forms with the RNA polymerase on glycerol gradient centrifugation, while the other cross-reacting proteins were not. Unambiguous evidence was obtained which indicated that the intracellular level of σ32 increased rapidly upon heatshock, at least in the strain containing high copy numbers of the rpoH gene.
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  • 22
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    Molecular genetics and genomics 210 (1987), S. 60-68 
    ISSN: 1617-4623
    Keywords: mRNA secondary structure ; Nuclease sensitivity ; Translational repressor ; Ribosomal protein ; Escherichia coli
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The secondary structure of the autoregulatory mRNA binding site of Escherichia coli ribosomal protein L1 has been studies using enzymatic methods. The control region of the E. coli L11 operon was cloned into a vector under control of the Salmonella phage SP6 promoter, and RNA transcribed using SP6 RNA polymerase. The secondary structure of this RNA was probed using structure-specific nucleases, and by comparison of the data with computer predictions of RNA folding, secondary structural features were deduced. The proposed model is consistent with elements of some previously proposed models, but differs in other features. Finally, secondary structure information was obtained from two mutant mRNAs and the structural features correlated with observed phenotypes of the mutants.
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  • 23
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    Molecular genetics and genomics 210 (1987), S. 364-368 
    ISSN: 1617-4623
    Keywords: Promoter region of the putA gene ; putC ; Nucleotide sequence ; Escherichia coli ; nitrogen control
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The nucleotide sequence of the putC region, from which divergent transcription of the putP and putA genes starts, was determined. The promoter region for the putA gene was restricted to the location between the HindIII site and the NcoI site or at the NcoI site by using putA-lacZ fusion plasmids and the transcriptional start for the putA gene was identified in the region between the HindIII site and the NcoI site by S1 mapping. This region also contains a potential CAP binding site, a ribosome binding site, and a sequence that is highly homologous to argTr. five potential promoters (putPp1-putPp5) for the putP gene, which were separate from the promoter region for the putA gene, were indicated by S1 mapping analysis of the putP gene transcripts. We concluded that the putC region is 419 bp long and contains two independent sets of promoters, regulating the expression of putP and putA genes in opposite directions. In addition, this region was found to contain an open reading frame (orf) capable of encoding a polypeptide of 111 amino acids in overlapping fashion. But studies using an orf-lacZ fusion gene showed that this open reading frame was not expressed.
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  • 24
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    Bulletin of experimental biology and medicine 103 (1987), S. 94-97 
    ISSN: 1573-8221
    Keywords: Escherichia coli ; B subunit ; diarrhea ; antibodies
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    Topics: Biology , Medicine
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  • 25
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    Bulletin of experimental biology and medicine 103 (1987), S. 372-374 
    ISSN: 1573-8221
    Keywords: Escherichia coli ; SSB proteins ; mutants
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    Topics: Biology , Medicine
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  • 26
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    Molecular genetics and genomics 206 (1987), S. 181-184 
    ISSN: 1617-4623
    Keywords: Escherichia coli ; Bacteriophage λ ; DNA loops ; DNA repair ; Gene conversion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Unmethylated DNA heteroduplexes with a large single stranded loop in one strand have been prepared from separated strands of DNA from two different strains of bacteriophage λ, one of which has a ∼800 base pair IS1 insertion in the cI gene. The results of transfections with these heteroduplexes into wild-type and mismatch repair deficient bacteria indicate that such large non-homologies are not repaired by the Escherichia coli mismatch repair system. However, the results do suggest that some process can act to repair such large non-homologies in heteroduplex DNA. Transfections of a series of recombination and excision repair deficient mutants suggest that known excision or recombination repair systems of E. coli are not responsible for the repair. Repair of large non-homologies may play a role in gene conversion involving large insertion or deletion mutations.
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  • 27
    ISSN: 1617-4623
    Keywords: mutT mutator ; Escherichia coli ; A:T→C:G transversion ; Molecular cloning ; Nucleotide sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The Escherichia coli mutator gene mutT, which causes A:T→C:G transversion, was cloned in pBR 322. mutT + plasmids carry a 0.9 kb PvuII DNA fragment derived from the E. coli chromosome. Specific labelling of plasmid-encoded proteins by the maxicell method revealed that mutT codes for a polypeptide of about 15,000 daltons. The protein was overproduced when the mutT gene was placed under the control of the lac regulatory region on a multicopy runaway plasmid. The nucleotide sequence of the mutT gene was determined by the dideoxy method.
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  • 28
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    Molecular genetics and genomics 207 (1987), S. 302-305 
    ISSN: 1617-4623
    Keywords: Escherichia coli ; IHF protein ; Plasmid F ; Conjugal transfer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We present evidence that the Escherichia coli DNA binding protein, IHF, plays an important role in conjugal transfer of the plasmid F. Our results suggest that IHF exerts this effect by positively effecting transcription of the transfer (tra) operon of the plasmid.
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  • 29
    ISSN: 1617-4623
    Keywords: Escherichia coli ; Iron hydroxamate transport ; fhuC, fhuD genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The transport of Fe3+ into cells of Escherichia coli occurs via siderophores and the uptake through the outer membrane of three Fe3+-siderophore compounds containing hydroxamate residues requires three specific receptor proteins. In contrast, transport through the cytoplasmic membrane is catalysed by three common proteins encoded by the fhuB, fhuC and fhuD genes. The nucleotide sequence of a DNA fragment containing the fhuC and fhuD genes has been determined: the open reading frame of fhuC contains 795 nucleotides which encode a polypeptide with a molecular weight of 29 255 and the largest open reading frame of the fhuD region comprises 888 nucleotides. However, we propose that translation of fhuD initiates at the fourth potential start codon resulting in a polypeptide with a molecular weight of 28 282. Both proteins are moderately nonpolar and membrane-bound. They lack obvious signal sequences. Segments of the FhuC protein display strong homology to ATP-binding proteins, suggesting a function in Fe3+ uptake similar to the ATP-binding proteins of transport systems that depend on periplasmic proteins. This study completes the nucleotide sequence of the fhu operon which consists of the four genes fhuA fhuC fhuD fhuB arranged in this order on the E. coli chromosome and transcribed from fhuA to fhuB.
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  • 30
    ISSN: 1617-4623
    Keywords: ssb ; Single-stranded DNA binding protein ; SOS repair ; RecA protein ; Escherichia coli
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Induction of the SOS response in Escherichia coli results in an increase in the relative rate of synthesis of single-stranded DNA binding protein (SSB). In contrast to RecA protein, this increase is slow and does not lead to higher SSB levels. The significance of ssb induction to SOS repair is discussed.
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  • 31
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    Molecular genetics and genomics 210 (1987), S. 1-4 
    ISSN: 1617-4623
    Keywords: Stringent control ; RNA polymerase ; Escherichia coli
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The quantitative effect of ppGpp, the effector of stringent control, on various Escherichia coli promoters was measured in an in vitro mixed transcription system. This allowed us to determine, among these promoters, the hierarchy of promoters according to their ppGpp susceptibility. The strength of the stringent control signal, however, was found to be altered when the test promoters were transcribed by ppGpp-insensitive RNA polymerases from relaxed mutants of E. coli, which carry substitutions in the β subunit gene (rpoB). Thus, it was concluded that the activity of the stringent control signal depends on the nature of the RNA polymerase as well as that of the promoter.
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  • 32
    ISSN: 1617-4623
    Keywords: Replication terminus ; Escherichia coli ; Y-shaped molecule
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A 4.32 kb DNA fragment, on which the DNA replication terminus (terR) site of plasmid R 6K was located, was inserted into the unique EcoRI site of plasmid pUC9. To detect replication intermediate molecules with a replication fork halted at the terR site, a cell DNA extract was digested with EcoRI, electrophoresed through an agarose gel and stained with ethidium bromide. In addition to two major bands, one derived from vector DNA and the other from the ter insert fragment, two extra minor bands were detected. Following DNA-DNA hybridization and electron microscopic observation we concluded that the two minor bands corresponded to the two Y-shaped molecules, produced from the θ-shaped intermediate molecules by EcoRI digestion.
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  • 33
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    Journal of industrial microbiology and biotechnology 1 (1987), S. 335-340 
    ISSN: 1476-5535
    Keywords: Escherichia coli ; Outer membrane permeability ; Porin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary We describe the construction and analysis of an isogenic series ofEscherichia coli K12 strains that vary in their outer membrane permeability. They carry mutations that alter the amount and the type of porin present in the outer membrane. The permeability profiles of these strains suggest that both the amount and the type of porin present in the outer membrane affects permeability. Several of the strains carry mutations that extend the permeability of the outer membrane to include a variety of compounds that are normally excluded from entering the cell.
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  • 34
    ISSN: 1476-5535
    Keywords: Fermentation ; High cell density ; Escherichia coli ; RecombinantE. coli ; Atrial natriuretic factor (ANF) ; Fusion protein expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Studies are presented on the fermentation of recombinantEscherichia coli that express rat atrial natriuretic factor (ANF) as a fusion protein. Our objective was to achieve high cell density while maintaining ANF expression at the same level as observed in shake flasks. Improved fermentation conditions included: maintaining glucose concentrations at 1 g/l, using an enriched medium, adding concentrates of medium throughout the fermentation, and blending oxygen for adequate aeration. Cell densities of 12 g/l (dry weight) were achieved, which represented a 10-fold increase over non-improved conditions, while maintaining ANF levels at 7 mg/g of dry cell mass. When galactose was used as an initial carbon source or as a feed supplement, there was a 2-3-fold increase in the expression of ANF from these high-cell-density fermentations. The recombinant ANF was biologically active.
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  • 35
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    Journal of industrial microbiology and biotechnology 2 (1987), S. 87-95 
    ISSN: 1476-5535
    Keywords: Fermentation ; Process control ; Expression of recombinant proteins ; Escherichia coli ; Malaria vaccine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary A variety of feeding strategies have been described for attaining high cell densities in fed-batch fermentors. Although cell density is an important component in the produtivity of recombinant fermentations, it must be achievable with high product expression levels. Experiments were conducted to study the influence of fermentation feeding strategies on the production of a recombinant malaria antigen inEscherichia coli. C-source feeding profiles were calculated to maintain specific growth rates at 0.1, 0.2, 0.35, and 0.5 l/h prior to induction in defined and complex media using an exponential growth model. Fed-batch fermentations employing these feeding profiles effectively controlled the specific growth rates prior to induction. Antigen yields per dry cell weight did not vary with specific growth rate. Antigen yields from fed-batch fermentations achieving high cell densities were similar to batch fermentations achieving low cell densities. These results show that C-feeding policies can limit growth without reducing expression levels in some systems, and suggest applications in managing oxygen demand and catabolic by-product formation during process scale-up.
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  • 36
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    Journal of industrial microbiology and biotechnology 1 (1987), S. 377-381 
    ISSN: 1476-5535
    Keywords: Antimicrobial ; Metal salt ; Lanthanum ; Escherichia coli ; Staphylococcus aureus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Rare earth metal cations are antibacterially synergistic with anionic surfactants, yielding mixtures that have bactericidal activity against a variety of gram-positive and gram-negative bacteria at minimum concentrations ranging from 16 to 125 μg/ml. Uptake of surfactant byEscherichia coli increases in the presence of lanthanum, suggesting that the role of rare earth metal cations is to reduce the net negative surface charge on the bacteria, thereby increasing the affinity between the negatively charged surfactant and the bacterial surface.
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