Herrmann und Ulrike / Band 1 (German Edition)

Italian Viola, Giuseppe Nadotti, Piacenza, c. Certificate; Fritz Reuter and Sons, More Bradshaw" in the plate l.

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Gouache on paper, sheet More Gouache on paper, sheet size 19 x 24 in. Caucasian Brocaded Prayer Rug, last quarter 19th century, slight brocade losses , 4 ft. Oil on canvas, 37 x 25 in. Herrmann, the round stick stamped A. Herrmann" in pencil l.

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Attributed to Curt Herrmann German, In the FieldUnsigned, identified on a presentation plaque affixed to the frame. Oil on canvas, 23 x 18 in. Search Your search has returned 78 results. Sixteen Rug and Textile Catalogs. German Nickel Silver-mounted Violin Bow. German Silver-mounted Cello Bow. Thirty-nine Rug Books and Magazines.

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Five Oriental Rug Books. Nickel Mounted Violin Bow. Silver Mounted Violin Bow. The supernatant S20 was saved. The pellet P20 was washed twice with RIPA buffer to avoid the presence of cytosolic proteins in the P20 sample and then resuspended in the original volume of the initial sample. Cells were washed three times in 0.

The second antibody conjugated with Alexa was added to the cells. After the final washing three times in 0. The hiAP assay was performed in 96 well microtiter plates as follows: Slopes of blanks were subtracted and activity was calculated with a path length of 5 mm and a millimolar extinction coefficient of All optimization experiments were done in a Sixfors bioreactor Infors AG, Bottmingen at ml scale. Cells were grown in SPP-based medium with regulation of pH 7.

IA adopted the hiAP assays and performed together with JR all of the fermentation experiments and participated in manuscript drafting. UB cloned the constructs and transformed the ciliates and made most of the Western blots and participated in manuscript drafting. WR participated in the development of the hiAP assays. SL made the immunofluorescence analysis and images.

TW made the de-glycosylation assays, participated in Western blots, immunofluorescence analysis and the conceptual work and prepared the manuscript. All authors read and approved the final manuscript. Detailed scan of hiAP expressing T. Example A and B: Here we show two detailed scans through a fixed cell that expresses full-length hiAP.

The single images are made in 0. The first example is the cell that is already shown in Figure 3B and C white box, detailed image. The arrows clearly demonstrate the surface localization of recombinant hiAP. The second structure is not further characterized, nevertheless it is present in all analyzed cells.

Example C and D: Here two further detailed scans are shown that illustrate the surface display of recombinant hiAP in T. The second internal structure is not further characterized, nevertheless it is present in all analyzed cells. Comparison of Tetrahymena thermophila with established expression systems. Overview of established expression systems compared to Tetrahymena thermophila.

National Center for Biotechnology Information , U. Published online Jan Received Sep 28; Accepted Jan This article has been cited by other articles in PMC. Abstract Background Tetrahymena thermophila possesses many attributes that render it an attractive host for the expression of recombinant proteins. Results Functional and full length human intestinal alkaline phosphatase was expressed by T. Conclusions With the expression of hiAP, not only a protein of commercial interest could be produced, but also a reporter enzyme that offers the possibility to analyze T.

Background The ciliate Tetrahymena thermophila is a small non pathogenic protozoan and one of the best characterized eukaryotic unicellular organisms. Results Expression and localization of human intestinal alkaline phosphatase in T. Open in a separate window. Secretion of functional hiAP into the medium In the next experiments we used an expression cassette encoding a hiAP truncation mutant that lacks the human GPI anchor signal to address two points. Optimization and monitoring of secretion in T. Discussion Here we report the expression of human alkaline phosphatase by the ciliate T.

Conclusions The precursor of the human intestinal alkaline phosphatase becomes correctly processed and is expressed as a functional enzyme. Methods Constructs To ensure gene expression in T. Strains, cell culture and transformation of T. Cell fractionation For cell fractionation experiments, 1 to 2 ml of T. AP activity assay The hiAP assay was performed in 96 well microtiter plates as follows: Optimization and fermentation All optimization experiments were done in a Sixfors bioreactor Infors AG, Bottmingen at ml scale.

Authors' contributions IA adopted the hiAP assays and performed together with JR all of the fermentation experiments and participated in manuscript drafting. Supplementary Material Additional file 1: Click here for file 21K, DOC. Isolation of micro- and macronuclei of Tetrahymena pyriformis. Beginning to understand the end of the chromosome. Continuous high-cell-density fermentation of the ciliated protozoon Tetrahymena in a perfused bioreactor. Mass cultivation of Tetrahymena thermophila yielding high cell densities and short generation times. Cultivation of Tetrahymena thermophila in a 1.

The circumsporozoite protein of Plasmodium falciparum is expressed and localized to the cell surface in the free-living ciliate Tetrahymena thermophila. Surface display of a parasite antigen in the ciliate Tetrahymena thermophila. Secretion of functional human enzymes by Tetrahymena thermophila. Parameters affecting the maximum cell concentration of Tetrahymena. The human alkaline phosphatases: Nucleotide and amino acid sequences of human intestinal alkaline phosphatase: Cloning and sequencing of human intestinal alkaline phosphatase cDNA.

Codon usage in Tetrahymena thermophila. Cloning and characterization of the gene encoding the highly expressed ribosomal protein l3 of the ciliated protozoan Tetrahymena thermophila. Evidence for differential codon usage in highly expressed genes.


High frequency vector-mediated transformation and gene replacement in Tetrahymena. Electroporation-mediated replacement of a positively and negatively selectable beta-tubulin gene in Tetrahymena thermophila.

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Herrmann und Ulrike / Band 1 (German Edition) eBook: Johann Karl Wezel: domaine-solitude.com: Kindle Store. Herrmann und Ulrike / Band 4 (TREDITION CLASSICS) (German Edition) [ Johann Karl Wezel] on domaine-solitude.com *FREE* shipping on qualifying offers. Dieses.

A robust inducible-repressible promoter greatly facilitates gene knockouts, conditional expression, and overexpression of homologous and heterologous genes in Tetrahymena thermophila. Secretion heterogeneity of lysosomal enzymes in Tetrahymena pyriformis. Biosynthesis of secreted beta-hexosaminidase in Tetrahymena thermophila. A comparison of the wild type with a secretory mutant. Three pools of lysosomal enzymes in Tetrahymena thermophila. Lysosomal enzymes produced by immobilized Tetrahymena thermophila. Secretion of acid hydrolases and its intracellular source in Tetrahymena pyriformis.

Genetic characterization of the secretory mutant MS-1 of Tetrahymena thermophila: Isolation and characterization of a mutant of Tetrahymena thermophila blocked in secretion of lysosomal enzymes. Cell biology of Tetrahymena thermophila.

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Out with a bang! Tetrahymena as a model system to study secretory granule biogenesis. Cloning and sequencing of four new metallothionein genes from Tetrahymena thermophila and T.

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Little, if any, attention has been paid to determine cultivation parameters that enhance the productivity of recombinant proteins. The double band probably corresponds to intracellular precursor hiAP that is membrane attached by the signal peptide or corresponds to a non-cleaved GPI anchor signal. Of course on the one hand this observation emphasizes that truncated hiAP provides a powerful tool to quantify the secretion efficiency in living ciliate cells by a very sensitive assay. To monitor the functionality of the human GPI anchor signal in the ciliate T. Kindle Edition File Size:

Cadmium metallothionein gene of Tetrahymena pyriformis. Biochemical and ultrastructural data on Tetrahymena pyriformis treated with copper and cadmium. Tetrahymena metallothioneins fall into two discrete subfamilies. Removal of phosphate from lipid A as a strategy to detoxify lipopolysaccharide. A physiologic function for alkaline phosphatase: Alkaline phosphatase treatment improves renal function in severe sepsis or septic shock patients. Enabled Average Customer Review: Be the first to review this item Would you like to tell us about a lower price? Would you like to report this content as inappropriate?

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