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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Current genetics 21 (1992), S. 83-84 
    ISSN: 1432-0983
    Keywords: Yeast ; Transformation ; Thio compound ; Stationary phase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A fast yeast-transformation technique has been developed by adding thio compounds to alkali-ion based protocols and incubating at 45°C. This procedure is especially recommended for cells from stationary phase at a density up to 2.5×108 cells/ml. It involves only one step for the preparation and transformation of competent cells within 30 min. The yield was more than 104 transformants/μg plasmid DNA. This protocol is easy to scale up for many DNA samples and is also applicable for yeast cells from different types of storages.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-072X
    Keywords: Bacterium ; Phosphorylation ; Protein kinase ; Xanthomonas campestris pv. oryzae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protein phosphorylation was studied in Xanthomonas campestris pv. oryzae in vivo and in vitro. In vitro labelling showed that the protein kinases in this bacterium used both ATP and GTP as nucleotide substrates at nearly the same efficiency. At least 6 proteins were phosphorylated in vitro, including abundant species of p81, p44, and p32 with M r of 81000, 44000, and 32000, respectively. Three types of phosphate-protein linkage were found in this bacterium: O-phosphate, N-phosphate and probably acyl phosphate. The p81 and p32 were phosphorylated at histidine. The p44 had mainly phosphoserine and a small part of phosphohistidine. The phosphorylation profile was variable depending on the growth conditions. Furthermore, by a virulent phage Xp10 infection the quantity of phosphorylation increased: for phosphohistinine more than 10-fold, and for phosphoserine about 3-fold. Thus, in this bacterium phosphorylation may be linked with a physiological regulation system and with Xp10 phage development.
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  • 3
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The σ subunit of Xanthomonas oryzae pv. oryzae is disassociated from host RNA polymerase after phage Xp10 infection. To clarify the possible mechanism for this observation, σ subunit was purified and an antiserum against σ subunit was prepared. Immunoprecipitation of RNA polymerase by the anti-core RNA polymerase antiserum, followed by immunoblotting with anti-σ subunit antibody, revealed that σ subunit was lost from RNA polymerase within 10 minutes after Xp10 infection. Loss of σ subunit was not observed under other stress conditions including heat and cold stress, starvation and growth to stationary phase. Two-dimensional immunoblotting analysis did not reveal any covalent modification of either σ subunit or RNA polymerase after Xp10 infection. These results suggest that separation of σ subunit from RNA polymerase may be due to competition with other binding factors.
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  • 4
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract Exposure of Xanthomonas oryzae pv. oryzae cells to 254 nm UV radiation resulted in an alteration of protein phosphorylation. Labelling of the phosphohistidine-containing proteins with molecular masses of 81 and 32 kDa, named p81 and p32, was rapidly reduced following UV irradiation in the early exponential cells, but the decrease was not detected in mid-exponential cells. Mitomycin C, a DNA replication inhibitor, and rifampicin, a drug generally used to inhibit RNA synthesis and DNA replication, were also found to reduce the histidyl phosphorylation. However, this alteration of protein phosphorylation was not hindered by chloramphenicol treatment. A possible role for these histidyl phosphopfoteins in sensing UV light is proposed.
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