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Immunocytochemical analysis of embryonic compartmentation with a monoclonal antibody against a cytokeratin-related antigen (1990)

Grunwald, G. B. ; Gilbert, S. F. ; Brewer, K. ; [et al.]

Springer

Histochemistry and cell biology 94 (1990), S. 545-553

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ISSN: 1432-119X

Source: Springer Online Journal Archives 1860-2000

Topics: Biology , Medicine

Notes: Summary Mab 113F4, a monoclonal antibody recognizing an antigen in the outer synaptic layer of the chick neural retina, also recognizes an antigen appearing in all three germ layers of the gastrulating chick embryo. However, as neurulation proceeds, the antigen is down-regulated in three distinct patterns. First, the antigen is lost specifically from those trunk ectodermal cells destined to form the neural plate and, later, the neural tube. It remains absent from any neural derivative until day 13 when it appears in the outer synaptic layer of the neural retina, coincident with synaptogenesis in this region. Second, the entirety of the head ectoderm loses this antigen as the head lifts off the blastoderm. This down-regulation is followed later by a similar loss of antigen expression in the trunk ectoderm. Third, expression in the mesoderm becomes limited to the lateral plate and extraembryonic epithelia. Endodermal derivatives continue to express the antigen throughout development. Antigen 113F4 is localized within the cytoplasm and is organized in a fibrillar pattern. The intracellular localization of this antigen and its characteristic spatio-temporal tissue distribution are consistent with the antigen being a cytokeratin or cytokeratin-related antigen. The changes in tissue distribution suggest a possible role in tissue modelling in response to inductive interactions during development.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/BF00272620

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Increase in copy number of an integrated vector during continuous culture of Hansenula polymorpha expressing functional human haemoglobin (1994)

Gilbert, S. C. ; van Urk, H. ; Greenfield, A. J. ; [et al.]

New York, NY [u.a.] : Wiley-Blackwell

Yeast 10 (1994), S. 1569-1580

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ISSN: 0749-503X

Keywords: Hansenula ; haemoglobin ; integration ; continuous culture ; Life and Medical Sciences ; Genetics

Source: Wiley InterScience Backfile Collection 1832-2000

Topics: Biology

Notes: Recombinant human haemoglobin A (rHbA) was produced by a leucine-requiring strain of Hansenula polymorpha which had been transformed with an integration vector containing the Saccharomyces cerevisiae LEU2 gene and cDNAs for the expression of α and β globin each driven by the H. polymorpha MOX promoter. After 40 generations in a chemostat it was found that the integrated vector had become amplified in the host strain. In some cases this led to an increase in LEU2 gene dosage, but a loss of globin expression cassettes. In other cases the globin gene dosage also increased. These changes coincided with an increase in rHbA production in the culture, which was reversed when the dilution rate was increased. Isolates from a chemostat culture producing elevated levels of rHbA were grown in fed-batch fermentations, resulting in higher productivities than when inoculated with the parent strain. The rHbA produced was purified and characterized. Oxygen binding studies and electrospray mass spectrometry showed that the rHbA had been processed and assembled correctly, and behaved as a fully functional co-operative tetramer.

Additional Material: 7 Ill.