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  • 1
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Bacterial pathogenesis is strictly regulated in response to changes in environmental conditions. A His–Asp phosphorelay system consisting of a sensor kinase and response regulator is used by Gram-positive and Gram-negative bacteria to control gene expression in response to environmental stimuli. We screened His–Asp phosphorelay systems for their effect on virulence expression in enterohaemorrhagic Escherichia coli (EHEC), and found rcsD or rcsB overexpression enhanced locus for enterocyte effacement (LEE) gene transcription and adherence to Caco-2 cells through transcriptional activation of the ler regulatory gene. An EHEC-specific regulator GrvA, encoded by ECs1274, was required for ler transcription activation by RcsB. Furthermore, GrvA activated ler transcription in E. coli K12. Stimulation of the RcsDCB regulatory system by RcsF overexpression slightly increased EspB expression in the wild type but not the ECs1274 mutant. However, EspB expression in an rcsB deletion mutant increased compared with wild type, suggesting that RcsB negatively regulates LEE gene expression and that active RcsB protein is present under normal growth conditions. Deletion of pchA, which encodes a positive regulator for ler, abolished the effect of the rcsB deletion, suggesting that pchA mediated the negative RcsB effect. pchA transcript levels decreased when RcsB expression increased. Thus, LEE gene transcription may be regulated by RcsB through two oppositely regulated O157-specific regulators, PchA and GrvA.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] FIG. 1 Northern hybridization of mRNAs from rat and human livers. Poly(A)* RNAs were isolated from livers of normal adult rat (lane a) and CCI4-treated rat (lane b). Lane c is human liver poly(A)+ RNA (Clontech). The positions of the 28S and 18S ribosomal RNA markers are shown on the left. Blots ...
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-041X
    Keywords: DNA injection ; CAT gene expression ; Midblastula transition ; Xenopus embryogenesis ; Coenocytic egg cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary InXenopus laevis embryogenesis, the synthesis of heterogeneous mRNA-like RNA starts at the cleavage stage, whereas that of low-molecular-mass RNAs and rRNA occurs at the early blastula and late blastula stages, respectively. In coenocytic fertilizedXenopus egg cells, which fail to cleave, an excess of exogenously injected DNA (pBR322) induces ‘premature’ expression of previously injected exogenous genes (yeast tRNA genes). We have carried out experiments to discover whether the injection of excess exogenous DNAs of various origins modifies the expression of endogenous genes and previously injected exogenous genes inXenopus embryogenesis. We found that injection of a relatively large amount of exogenous DNA (Xenopus rDNA clone) induces the premature expression, or enhanced expression, of previously injected bacterial chloramphenicol acetyltransferase genes in coenocytic cells. In embryos, however, the injection of exogenous DNAs of various origins did not appreciably modify the expression of either endogenous or previously injected CAT genes. The DNAs injected into fertilized eggs were not degraded and were partitioned into the nuclei of most (at least 80%) of the descendant blastomeres at least during early stages of development. Therefore, we concluded that the program of gene expression in normally developing embryos cannot easily be altered by the introduction of excess exogenous DNAs.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Development genes and evolution 205 (1996), S. 300-310 
    ISSN: 1432-041X
    Keywords: Xenopus laevis HGF gene promoter ; CAT fusion gene ; Temporally and spatially controlled expression ; Silencer elements ; Mesoderm induction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Previously, we isolated Xenopus HGF (hepatocyte growth factor) cDNA and showed in Xenopus embryos that expression of this gene starts at the late gastrula stage mainly in the ventral mesoderm, and furthermore that the expression is induced in animal cap by activin A and bFGF (basic fibroblast growth factor). Here we have cloned the Xenopus HGF gene, covering a 14 kb 5′-upstream region and a 0.2 kb 5′-coding region. Within about 0.5 kb of the 5′-flanking region, the Xenopus HGF gene contained a TATA-like element AATGAAA, one putative NF-1 binding site, two NF-IL-6 binding motif sequences, one putative TGF-β-dependent inhibitory element (TIE) and one AP-1 binding site. A recombinant circular plasmid consisting of a 1.7 kb HGF promoter region and the bacterial chloramphenicol acetyltransferase (CAT) gene was first expressed at the late gastrula stage in the ventral mesoderm, as was the endogenous HGF gene. The expression of the fusion gene was induced in animal cap cells by activin A and bFGF although induction by the latter was not so strong. Using a series of 5′-deletion constructs introduced into animal caps, silencer elements, which seem to be essential for the gene's regionally correct expression, and the element responsible for induction by activin were found. The results show that the HGF gene promoter isolated here contains elements which may endow the gene with the regulative function for its temporally and spatially regulated expression, although the element necessary for induction by bFGF seems to be missing.
    Type of Medium: Electronic Resource
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