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  • 1
    ISSN: 0749-503X
    Keywords: LFH deletion cassette ; functional analysis ; chromosome IV ; Saccharomyces cerevisiae ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We report here the construction of six deletion mutants and the analysis of their basic phenotype. Deletion cassettes containing the KanMX4 marker module and long flanking regions homologous to the target locus were constructed for each of the six open reading-frames (ORFs YDL088c, YDL087c, YDL086w, YDL085w, YDL084w and YDL082w) located on chromosome IV. Sporulation and tetrad analysis of heterozygous deletant strains revealed that, in the FY1679 genetic background, ORFs YDL088c, YDL087c and YDL084w are essential genes for vegetative growth whereas YDL086w, YDL085w and YDL082w are non-essential. ydl088cΔ and ydl084wΔ haploid strains are viable in the CEN. PK2 genetic background although ydl084wΔ grows at a slower rate than the wild type. Complementation tests by corresponding cognate genes confirmed that gene inactivation was responsible for these growth defects. © 1998 John Wiley & Sons, Ltd.
    Additional Material: 1 Ill.
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  • 2
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome VII ; ribonuclease PH ; HGH1 ; YGR187c ; YGR189c ; YGR194c ; YGR195w ; YGR196c ; YGR198w ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have deleted six different ORFs of unknown function located on the right arm of Saccharomyces cerevisiae chromosome VII; namely, YGR187c/HGH1, YGR189c, YGR194c, YGR195w, YGR196c and YGR198w. No basic phenotypes could be attributed to the strains deleted in any of genes YGR187c/HGH1, YGR189c, YGR194c and YGR196c. These deletants did not show mating, sporulation or growth defects under any of the conditions tested. However, spores bearing deletions in either the YGR195w or YGR198w genes were unable to develop into macroscopical colonies. The YGR195w gene product shows significant homology with bacterial ribonuclease PH, an enzyme hitherto undescribed in yeasts, and its deletion causes a loss of viability after one to three rounds of cell division. Overexpression of this gene, using a tetracycline-regulatable promoter system, did not cause any effect on the cells. Contrary to what has been reported for prokaryotic homologs, this enzyme could play an essential role in yeast cell biology. The product encoded by the other essential ORF, YGR198w, shows no significant homology with any protein of known function in the databases. Spores bearing the deletion usually germinate and give rise to microcolonies of 50-100 non-viable cells. © 1998 John Wiley & Sons, Ltd.
    Additional Material: 2 Ill.
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  • 3
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Electrophoresis 19 (1998), S. 1117-1126 
    ISSN: 0173-0835
    Keywords: Purification ; Free-flow electrophoresis ; Transport phenomena ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Free-flow zone electrophoresis may be used to purify biological samples, due to differences in electrophoretic mobility, in absence of a matrix - most frequently a gel - thus enabling the biological integrity of even fragile molecules to be preserved. However, the process is more complicated than its principle suggests due to different transport phenomena interfering with electrophoretic migration, with the resultant separation depending both on separation effects and dispersive phenomena. The physical origin of the main effects involved was identified. Mathematical expressions were proposed to estimate the influence of the crescent effect and electrohydrodynamics on the process. In this paper, these equations are used to determine the minimum difference in electrophoretic mobility required for a separation to be achieved with respect to the processing parameters. A methodology is proposed which defines the conditions under which the difference in electrophoretic mobilities equals that calculated when considering the influence of dispersive phenomena. Optimized separations of the whey proteins lactoferrin and albumin, known to interact strongly, and the purification of a monoclonal antibody from a mouse ascitic fluid illustrate the approach.
    Additional Material: 15 Ill.
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  • 4
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Electrophoresis 19 (1998), S. 1294-1299 
    ISSN: 0173-0835
    Keywords: Isoelectric focusing ; Continuous free flow electrophoresis ; Purification ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: To estimate the feasibility of isoelectric focusing in a continuous mode, the conditions for obtaining stable and linear pH gradients were first investigated, using a mixture of carrier ampholytes. Then the operating parameters allowing the isoelectric focusing of bovine serum albumin as model protein were determined. A theoretical approach was used to find conditions minimizing the electrohydrodynamic effect, mainly responsible for protein dispersion, for the separation of bovine serum albumin and α-lactalbumin from bovine milk.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
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