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  • 1
    ISSN: 1546-1696
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: [Auszug] Infection of the mammary gland, in addition to causing animal distress, is a major economic burden of the dairy industry. Staphylococcus aureus is the major contagious mastitis pathogen, accounting for approximately 15–30% of infections, and has proved difficult to control using standard ...
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  • 2
    ISSN: 1546-1696
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: [Auszug] Uroplakin genes are expressed in a bladder-specific and differentiation-dependent fashion. Using a 3.6-kb promoter of mouse uroplakin II gene, we have generated transgenic mice that express human growth hormone (hGH) in their bladder epithelium, resulting in its secretion into the urine at ...
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  • 3
    ISSN: 1546-1696
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: [Auszug] Mastitis, the most consequential disease in dairy cattle, costs the US dairy industry billions of dollars annually. To test the feasibility of protecting animals through genetic engineering, transgenic cows secreting lysostaphin at concentrations ranging from 0.9 to 14 mg/ml in their milk were ...
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  • 4
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Company
    Nature biotechnology 15 (1997), S. 416-417 
    ISSN: 1546-1696
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: [Auszug] Genetically engineered livestock have major advantages over transgenic mice for many purposes. Because of their size, relatively long life span, and physiological similarities to humans, livestock are ideally suited for use as genetic disease models. Furthermore, they are being employed as ...
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  • 5
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] Studies with mouse ova indicated that integration of a gene into host chromosomes is much more efficient with nuclear than with cytoplasmic injection14. On this basis, we reasoned that nuclear injection would be an appropriate first approach with other species. The first problem encountered was ...
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Transgenic research 8 (1999), S. 313-315 
    ISSN: 1573-9368
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
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  • 7
    ISSN: 1573-9368
    Keywords: codon bias ; IRES ; MAR ; rtTA ; TetR ; tetR ; transgenic ; transgene switch
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The objective of this work was to further develop a tetracycline repressor (TetR) protein system that allows control of transgene expression. First, to circumvent the need for a binary approach, a single plasmid design was constructed and tested in tissue culture. To indirectly assay integrations that express the synthetic transcription factor (rtTA), a bicistronic gene was built which included an internal ribosome entry site (IRES) and a green fluorescent protein coding region (GFP) on the same expression cassette as the coding region of rtTA (pTetGREEN). This construct did not produce fluorescent colonies when stably integrated and provided minimal expression of GFP in the face of adequate expression of rtTA. The coding region for TetR was then altered by introducing 156 silent point mutations to simulate mammalian genes. Replacement of wild‐type TetR gene (tetR) in pTetGREEN with 'mammalianized' provided GFP expression. Adjustment of codon usage in the tetR region of rtTA nearly doubled the expression level of functional rtTA. To increase the number of rtTA expressing lines, the chicken egg‐white lysozyme matrix attachment region (MAR) was introduced into the single plasmid design just upstream of the tetracycline operators (tetO). Inclusion of the MAR doubled the number of colonies that expressed rtTA (44 vs 88). With the modifications described here, the number of lines that express rtTA and provide induction from a single plasmid design can be increased by the inclusion of a MAR and the level of rtTA expression can be further increased by adjusting the base composition of the TetR coding region. The MAR also insulates the inducible gene from the promoter driving rtTA.
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Transgenic research 4 (1995), S. 39-43 
    ISSN: 1573-9368
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In general, genomic transgenes are expressed efficiently in mice, while their cDNA-based transgenes are frequently silent. Clarket al. (1992) have shown that silent cDNA transgenes under the control of the sheep β-lactoglobulin promoter can be activated after co-injecting them with a genomic sheep β-lactoglobulin transgene. We have tested the general utility of this concept using mouse whey acidic protein (WAP) transgenes. Here we show that WAP cDNA transgenes are virtually silent in transgenic mice. In contrast, WAP transgenes containing all introns are expressed in approximately 50% of the lines at levels ranging from 1% to more than 100% of the endogenous RNA (McKnightet al., 1992). When a WAP-genomic transgene was co-injected with a WAP-cDNA, basal activation of the cDNA was possible. However, the activity of the WAP-cDNA transgene did not exceed 1% of the WAP-genomic transgene. This suggests that a permissive integration site capable of supporting basal level transcription can be established, but further events are required for full activation of the transgene.
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  • 9
    ISSN: 1573-9368
    Keywords: whey acidic protein gene ; transgenic sheep ; bioreactor ; mammary gland
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The synthesis of foreign proteins can be targeted to the mammary gland of transgenic animals, thus permitting commercial purification of otherwise unavailable proteins from milk. Genetic regulatory elements from the mouse whey acidic protein (WAP) gene have been used successfully to direct expression of transgenes to the mammary gland of mice, goats and pigs. To extend the practical usefulness of WAP promoter-driven fusion genes and further characterize WAP expression in heterologous species, we introduced a 6.8 kb DNA fragment containing the genomic form of the mouse WAP gene into sheep zygotes. Two lines of transgenic sheep were produced. The transgene was expressed in mammary tissue of both lines and intact WAP was secreted into milk at concentrations estimated to range from 100 to 500 mg/litre. Ectopic WAP gene expression was found in salivary gland, spleen, liver, lung, heart muscle, kidney and bone marrow of one founder ewe. WAP RNA was not detected in skeletal muscle and intestine. These data suggest that unlike pigs, sheep may possess nuclear factors in a variety of tissues that interact with WAP regulatory sequences. Though the data presented are based on only two lines, these findings suggest WAP regulatory sequences may not be suitable as control elements for transgenes in sheep bioreactors.
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  • 10
    ISSN: 0730-2312
    Keywords: tetracycline ; MMTV-LTR ; tTA ; MMTV-tTA ; β-galactosidase ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Molecular mechanisms of development and disease can be studied in transgenic animals. Controlling the spatial and temporal expression patterns of transgenes, however, is a prerequisite for the elucidation of gene function in the whole organism. Previously we reportted that mice carrying a tetR/VP16 hybrid gene (tTA), under the control of the hunman cytomegalovirus immediate early 1 (HCMV-IE1) gene promoter, can be used to temporally activate the expression of transgenes under the control of a promoter containing tetop sequences. We now show that the MMTV-LTR can be used to target expression of tTA to the epthelial cells of secretory organs and skin in transgenic mice. Notably, nearly uniform expression of a tetop-lacZ transgene was found in seminal vesicle, salivary gland, and Leydig cells of mice carrying also the MMTV-tTA transgene. More heterogeneous patterns of gene expression were observed in mammary epithelial cells and nasal cells of the epidermis. Different MMTV-tTA lines had conparable tissue expression patterns. Transcriptional activation mediated by tTA was up to several hundred fold and it was abrogated after the administration of tetracycline. The MMTV-tTA mice established in this work will be useful for experiments examining the roles of biological factors at defined developmental stages in the epithelial cells of salivary gland, seminal vesicle, mammary gland, and skin and the Leydig cells of testes in addition, in combination with the CRE/lox recombnation system, these mice will be useful to achieve gene deletions at defined time points in these organs. © 1995 Wiley-Liss, Inc.
    Additional Material: 2 Ill.
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