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  • 1
    Electronic Resource
    Electronic Resource
    The 40-kilodalton allergen of Candida albicans is an alcohol dehydrogenase: molecular cloning and immunological analysis using monoclonal antibodies (1991)
    Oxford, UK : Blackwell Publishing Ltd
    Clinical & experimental allergy 21 (1991), S. 0 
    Details
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: To characterize the 40-kilodalton (kD) major allergen of Candida albicans (C. albicans), six monoclonal antibodies (MoAbs) against this allergen were generated. In SDS-polyacrylamide gel electrophoresis and immunoblot analysis, these MoAbs showed four different reaction patterns to antigens of six different Candida species. With the exception of one MoAb, other MoAbs were resistant to periodate treatment indicating non-carbohydrate epitopes were probably being recognized by these MoAbs. These MoAbs were used in the molecular cloning and immunological analysis of the gene coding for the 40-kD allergen. Nucleotide sequence determination of the two λgtl 1 cDNA clones obtained showed that the 40-kD allergen is an alcohol dehydrogenase (ADH) which shares a 70% amino acid sequence homology with the ADH isozyme I of Saccharomyces cerevisiae. This finding was confirmed by positive immunological response of the lysates of the clones obtained and a preparation of ADH of Saccharomyces cerevisiae to various MoAbs and to IgE antibodies in sera of allergic patients.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1365-2222.1991.tb03195.x
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  • 2
    Electronic Resource
    Electronic Resource
    Biogenesis of mitochondria 47 (1977)
    Springer
    Molecular genetics and genomics 153 (1977), S. 279-288 
    Details
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary This paper consolidates and refines the physical map of genetic loci previously established in our laboratory, by molecular analysis of seven genetically characterized new petites (deletion mutants of mtDNA). A modified DNA-DNA hybridization procedure employing filters simultaneously bound with mtDNA from two different petites has been used to measure the overlaps in mtDNA sequences between the different petite mutants. Thus, by analysis of three new petites carrying the antibiotic-resistance loci, ery1, cap1 and par1 on their mitochondrial genomes, it has now been possible to improve our estimation of the maximum distance between the cap1 and ery1 loci. The cap1, ery1 loci, and the 21S ribosomal RNA gene have now been mapped within 5 units in the same region (map position 0 to 5 units). Similarly, by analysis of four new petites carrying the O II and/or par1 loci on their mtDNAs, the map position of the O II locus is also more accurately determined within 2 units in a region (map position 34 to 36 units) between the par1 and ana1 loci. The positions of other loci including par1, the 15S ribosomal RNA gene, and some mit - loci are also discussed. We have thus extended our library of genetically and molecularly defined petite mutants, resulting in a set of petites having overlapping regions distributed throughout the entire wild-type mitochondrial genome, consistent with the idea that yeast mtDNA is physically circular.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00431593
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  • 3
    Electronic Resource
    Electronic Resource
    Biogenesis of mitochondria. 53 (1979)
    Springer
    Molecular genetics and genomics 174 (1979), S. 307-316 
    Details
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The mitochondrial DNA (mtDNA) of sevcral petite mutants of Saccharomyces cerevisiae characterized for the loss or retention of genetic loci in the region of the 21S rRNA gene has been analysed. RNA-DNA hybridization has indicated the presence of deletions within the 21S rRNA gene of some of these petites. The positions of the deletions were mapped precisely using the restriction enzymes HpaII, HaeIII and SalI. The results permitted the determination of the restriction sites relative to the 21S rRNA gene. By comparing the retention and loss of genetic loci with the mtDNA sequences present in each petite, physical map positions for the ery1, tsr1 and ω loci were assigned within the 21S rRNA gene. However, the data for the cap1 locus did not permit an unambiguous assignment of this locus inside or outside of the 21S rRNA gene. In some petites deleted for the tsr1 locus the behaviour of the ω locus underwent a change from {ie307-1}.to {ie307-2}, and this was shown to be accompanied by a deletion of 1.5 Kbp from within the 21S rRNA gene. The interference from this observation is that {ie307-3} behaviour can arise by deletion of sequences from {ie307-4}. This contrasts with the previously published observations that {ie307-5} strains contain an inserted sequence of about 1 Kbp that is not present in {ie307-6} strains. The rationalization of this apparent contradiction is that ω behaviour is not necessarily determined by a particular short nucleotide sequence, but rather, ω action reflects in some way the sum total of sequences retained or lost in this region of mtDNA.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00267804
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  • 4
    Electronic Resource
    Electronic Resource
    Controlled fed-batch fermentation of recombinant Saccharomyces cerevisiae to produce hepatitis B surface antigen (1988)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 32 (1988), S. 334-340 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: We have performed controlled fed-batch fermentation experiments to compare the production level of hepatitis B surface antigen (HBsAg) by recombinant yeast Saccharomyces cerevisiae strains (YNN27/pYBH-1, YNN27/ p2μ-S11, YNN27/pDCB-S2) containing plasmid vector with alcohol dehydrogenase (ADH1), acid phosphatase (PHO5), and glyceraldehyde-3-phosphate dehydrogenase (GPD) promoter, respectively. Yeast cell concentrations of 15-35 g dry cell weight/L were obtained. By limiting phosphorous concentration, HBsAg expression level for the YNN27/p2μ-S11 strain with inducible PHO5 promoter reached 0.2-0.3 mg/L. By controlling nutrient addition rate and dissolved oxygen concentration, HBsAg concentrations of 3-10 mg/L were achieved in 60-70 h fermentation using the YNN27/pDCB-S2 strain with the constitutive GPD promoter.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260320310
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  • 5
    Electronic Resource
    Electronic Resource
    Frequent deletions and sequence aberrations at the transgene junctions of transgenic mice carrying the papillomavirus regulatory and the SV40 TAg gene sequences (1995)
    Springer
    Transgenic research 4 (1995), S. 52-59 
    Details
    ISSN: 1573-9368
    Keywords: transgenic mice ; transgene junctions ; DNA integration ; sequence deletion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Exogenous DNA microinjected into one-cell mouse zygotes either integrates into the host genome within a short time span, or is rapidly degraded. On integration, a transgene squence is frequently reiterated. In this report, we describe the enzymatic amplification analysis of transgene junctions of 12 transgenic mice carrying different copy numbers of the same transgene with dissimilar ends. The transgene was composed of the regulatory sequence of the type 18 human papillomavirus linked to the TAg gene of the SV40 virus. Nucleotide sequences of 36 of these junctions were also determined. Deletions were found in 33 (91.7%) of the junctions analysed. At the crossover regions, 55.6% contained short overlapping sequences of one to six nucleotides. Insertions of 2–6 extraneous nucleotides were also found in 8.3% of the transgene junctions. Within a 10-nucleotide sequence on both sides of the transgene junctions, topoisomerase I (topo I) cleavage sites, runs of homogeneous purines or pyrimidines, alternating purine-pyrimidine tracks and (A-T)-rich sequences were found frequently. Stringent control experiments were also performed to ascertain that the observations made were not artefacts resulting from the polymerase chain reaction. Our data therefore indicate that damage had occurred quite frequently and extensively in our transgene construct. Such transgene damage may also occur to various extents in mice carrying other transgenes. Primary structure of the nucleotide sequences of the injected DNA seems to influence the process of transgene reiteration and aberration.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01976502
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  • 6
    Electronic Resource
    Electronic Resource
    An improved scheme for the identification of antigens recognized by specific antibodies in two-dimensional gel electrophoresis and immunoblotting (1990)
    Weinheim : Wiley-Blackwell
    Electrophoresis 11 (1990), S. 878-882 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: This paper describes an improved scheme for the identification of antigens in crude extracts recognized by specific antibodies when analyzed by a combination of two-dimensional gel electrophoresis and immunoblotting. First, protein components in gels are electrophoretically transferred to a polyvinylidene difluoride membrane which does not shrink or change dimensions in organic solvents. The efficiency of transfer and the localization of sample proteins on the membrane are checked and recorded by staining the blotting membrane with Fast Green FCF and recording the profile on a transparency. After blocking and the immunoassay, the results are recorded by photography. The sites of immune reaction are marked and the same membrane is restained briefly with Coomassie Brilliant Blue R-250 for the protein profile. Thus antigens in complex mixtures, recognized by antibodies of interest, can easily be identified from the restained membrane. If the whole protein profile is not well demonstrated, when used in combination with the profile recorded on the transparency, spots appearing on the restained membrane can still be used as useful landmarks in the final unequivocal antigenic identification. This improved scheme circumvents problems arising from membrane shrinkage and difficulties in accurately matching immunoreactive spots by conventional procedures and thus provides an accurate, simple and fast approach in the identification of antigens after immunoblotting.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150111018
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