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  • 1
    ISSN: 1546-1696
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: [Auszug] Transgenic livestock may prove useful for the large scale production of valuable proteins. By targeting expression to the mammary gland these proteins could be harvested from milk. To this end, we have designed a hybrid gene to direct the synthesis of human anti-hemophilic factor IX to the mammary ...
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1546-1696
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: [Auszug] It is over ten years since the birth of Dolly was announced amid enormous media attention. Today, the cloning of livestock animals is being pursued with a wide range of applications in mind, such as improved breeding efficiencies, enhanced and enriched food traits, the preservation of endangered ...
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Cellular and molecular life sciences 47 (1991), S. 905-912 
    ISSN: 1420-9071
    Keywords: Gene transfer ; gene modification ; gene expression ; livestock ; transgenic animal ; pharmaceutical proteins ; milk composition
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract There is every reason to expect that it will be possible within the next few years to begin to use farm animals to produce large quantities of some of the human proteins that are needed for the treatment of disease. Revolutionary new opportunities for the production of novel proteins in milk have been created by the development of methods for gene transfer. Exploitation of these opportunities depends upon selection and cloning of milk protein genes and identification of the sequences that govern tissue specific hormonally induced expression in the mammary gland. Studies with three genes, ovine β-lactoglobulin, rat β-casein and whey acidic protein of rat and mouse, suggest that they may all meet this requirement. Fragments of the ovine β-lactoglobulin, murine whey acidic protein and rabbit β-casein genes have directed production of novel proteins in the milk of transgenic mice, sheep, rabbits and pigs. The proteins were biologically active and usually co-migrated with authentic proteins. In early experiments, protein concentration was low, but our recent observations suggest that fusion genes containing genomic clones direct production of concentrations of protein that are suitable for commercial exploitation. In the longer term, two approaches may offer the potential of more reliable expression. Control elements capable of directing expression that is independent of site of insertion of the gene, but dependent on the number of copies of the gene, have been identified for a small number of genes. The availability of such elements for the milk protein genes would increase the reliability of gene expression considerably. Alternatively, targeted mutation of genes may allow the insertion of coding sequences within an existing gene so avoiding position effects.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1617-4623
    Keywords: Key words Introns ; Mammary gland ; Ovine genome ; Splicing ; Transgenes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Eukaryotic pre-mRNAs undergo a variety of post-transcriptional modifications, including the removal of intronic sequences by splicing, leading to creation of a functional mRNA. We have compared the processing of transcripts generated from ovine β-lactoglobulin gene constructs in stably transfected cells and in transgenic mice. In both the in vitro and in vivo model systems the removal of the middle two introns resulted in the inefficient splicing of the downstream, terminal intron. This intron-containing transcript was detected in the cytoplasmic RNA fraction. Thus, the initial in vitro analysis in cell lines of minigene constructs destined for expression in transgenic animals may provide a rapid and reliable indicator of the processing efficiency of the pre-mRNA produced by the construct in vivo. This is in contrast to the apparent limitations of in vitro systems in the analysis of transcription regulatory elements required for transgene expression.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 252 (1996), S. 465-469 
    ISSN: 1617-4623
    Keywords: Introns ; Mammary gland ; Ovine genome ; Splicing ; Transgenes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Eukaryotic pre-mRNAs undergo a variety of post-transcriptional modifications, including the removal of intronic sequences by splicing, leading to creation of a functional mRNA. We have compared the processing of transcripts generated from ovineβ-lactoglobulin gene constructs in stably transfected cells and in transgenic mice. In both the in vitro and in vivo model systems the removal of the middle two introns resulted in the inefficient splicing of the downstream, terminal intron. This intron-containing transcript was detected in the cytoplasmic RNA fraction. Thus, the initial in vitro analysis in cell lines of minigene constructs destined for expression in transgenic animals may provide a rapid and reliable indicator of the processing efficiency of the pre-mRNA produced by the construct in vivo. This is in contrast to the apparent limitations of in vitro systems in the analysis of transcription regulatory elements required for transgene expression.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1617-4623
    Keywords: Key words Chromatin ; Mammary gland ; Ovine ; Transgene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The ovine milk protein β-lactoglobulin is expressed in a distinct temporal pattern during lactogenesis. This expression pattern is reflected in the temporal profile of appearance of DNase I hypersensitive sites (HS) associated with the β-lactoglobulin gene in the mammary gland. Specifically, HSIV and HSV are present prior to the first major increase in expression, which occurs at mid-pregnancy, while HSI displays the converse profile, being detected after mid-pregnancy and during lactation. The extent of DNase I digestion at HSIII, encompassing the promoter region, reflects the level of β-lactoglobulin expression. In transgenic mouse mammary chromatin, β-lactoglobulin transgenes display the same set of DNase I hypersensitive sites as in sheep mammary chromatin. The temporal profile, however, differs from that seen in sheep: notably, HSIV and HSV are detected during lactation. The fact that β-lactoglobulin transgenes lacking HSIV and HSV are expressed but display a reduced transcription rate per integrated copy is compatible with a functional role for these regions. This suggests that HSIV and HSV may increase the likelihood of high-level transgene expression.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Cytotechnology 31 (1999), S. 3-8 
    ISSN: 1573-0778
    Keywords: cell culture ; livestock ; milk ; nuclear transfer ; transgenic
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Transgenesis may allow the generation of farm animals with altered phenotype, animal models for research and animal bioreactors. Although such animals have been produced, the time and expense involved in generating transgenic livestock and then evaluating the transgene expression pattern is very restrictive. If questions about the ability and efficiency of expression could be asked solely in vitro rapid progress could be achieved. Unfortunately, experiments addressing transcriptional control in vitro have proved unreliable in their ability to indicate whether a transgene will be transcribed or not. However, initial studies suggest that cell culture may be able to predict in vivo post-transcriptional events. We review these issues and propose that strategies which engineer the transgene integration site could enhance the probability for efficient expression. This approach has now become feasible with the development of techniques allowing animals to be generated from somatic cells by nuclear transfer. The important step in this procedure is the use of cells grown in culture as the source of genetic information, allowing the selection of specific transgene integration events. This technology which has dramatically increased the potential use of transgenic livestock for both agricultural and biotechnological applications, is based on standard cell culture methodology. We are now at the start of a new era in large animal transgenics.
    Type of Medium: Electronic Resource
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