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  • Springer  (3)
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  • 1
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A variant of Saccharomyces cerevisiae pep4 strain 20B12, with improved oligotrophic proliferation, cell survival and secretion of heterologous mouse α-amylase, is described. Previously we reported a procedure to enrich NI transformants that are not inhibited by cytotoxic expression of hepatitis B virus surface antigen in the secretion pathway of the protease-A-deficient (pep4) strain. To use the NI cells as a host for heterologous expression, we tried to amend the introduced pYAS/12S vector and obtain a host strain, NI-C, with stable NI phenotype and trp1 marker restored. Southern analysis of genomic DNA of NI-C suggested that the original pYAS/12S was abnormally rearranged and not completely corrected. Further assay showed that the viability and mitotic ability of the NI-C strain were increased. While using the NI-C strain as host for plasmid transformation and heterologous expression of mouse α-amylase, we observed that transformed colonies grew more quickly and secreted more α-amylase than general yeast strains. A further test showed that the NI-C strain was able to use mouse α-amylase as a positive selection marker to form transformed colonies on nitrogen-starved plates that contain starch as the sole carbon source. The results imply that the NI-C variant is an improved pep4 strain that can be used for heterologous expression and for the development of new selective markers in the yeast transformation system.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract An efficient one-step transformation method for the dimorphic yeast Yarrowia lipolytica is described. Using cells grown overnight on agar plates, the whole process is carried out within 1 h. The transformant clones could be recovered on selective plates as early as 36–48 h after plating. The efficiency was better than 105 transformants/μg replicative plasmid DNA. Effects of cell density, dithiothreitol, heat shock, poly(ethylene glycol) 4000 concentration and the wetness of selective plates were investigated.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1619-7089
    Keywords: Gated blood pool ; Computer processing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We studied the application of linear discriminant analysis to the computer assisted interpretation of rest and exercise gated blood pool studies. Data were obtained in 45 patients in whom the presence or absence of coronary artery disease was determined by coronary angiography. Automated analysis of regional ejection fractions using a linear discriminant function (LDF) was compared to the fully automated calculation of global ejection fraction and to the subjective evaluation of wall motion. A LDF constructed from the data for the first 31 consecutive patients correctly classified 25/31 (81%). The change in global ejection fraction correctly classified 24/31 (77%) and wall motion analysis 25/31 (81%). In a distint group of 14 patients the same LDF correctly classified 12/14 (86%), global ejection fraction 9/14 (64%) and wall motion analysis 10/14 (71%).
    Type of Medium: Electronic Resource
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