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  • 1
    ISSN: 1573-9368
    Keywords: human lactoferrin ; αS1-casein ; transgenic mice ; tissue specificity ; mammary gland
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The expression of human lactoferrin (hLF) in the milk of transgenic mice is described. Regulatory sequences derived from the bovine αS1-casein gene were fused to the coding sequence of the hLF cDNA and several lines of transgenic mice were generated. Human LF RNA was detected exclusively in the mammary gland of lactating females and only after the onset of lactation. No aberrant RNA products could be detected using northern blotting and primer extension analysis. The hLF concentrations in the milk ranged from less than 0.1 to 36 μg ml−1. Human LF thus expressed did not differ from human milk derived LF, with respect to molecular mass and immunoreactivity with monoclonal and polyclonal antibodies.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-9368
    Keywords: transgenic ; mammary gland ; casein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The bovine αs1-casein gene, isolated from a cosmid library, was introduced into the murine germline. Transgene expression occurred in all transgenic mice, and was confined to the lactating mammary gland. Half of the mouse lines (five out of ten) expressed at relatively high expression levels (〉1 mg ml−1). The highest levels of expression were obtained with a transgene containing 14.2 kb of 5′ flanking sequence, in two cases expression levels comparable to (10 mg ml−1) or well above (20 mg ml−1) αs1-casein levels in bovine milk were obtained. Transcription initiation occurred at the same site in the bovine αs1-casein gene in transgenic mouse as in the cow. A marked induction of expression occurred at parturition rather than at mid-pregnancy, and thus resembled the bovine rather than the murine developmental expression pattern. Bovine αs1-casein specific immunoblotting and RIA were developed for characterization and quantificatio n of the recombinant protein. Using these assays, the properties of the recombinant protein could not be distinguished from those of the natural bovine protein. In spite of the high-level tissue-specific and correctly regulated developmental expression of the transgene, expression levels were integration-site dependent. This may indicate that not all cis-acting regulatory elements involved in bovine αs1-casein expression were included in the transgene
    Type of Medium: Electronic Resource
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