Maschio, Antonio; (1990) Expression of the guinea-pig alpha-lactalbumin gene in transgenic mice. Doctoral thesis (Ph.D), UCL (University College London).

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Abstract

In order to identify the mechanisms at work in the gene regulation of milk protein genes, and in particular the α-lactalbumin gene, the guinea pig α-lactalbumin gene has been analysed both In Vitro using a mobility shift assay and In Vivo by the creation of transgenic mice. Results obtained using the In Vitro system demonstrated the formation of specific complexes by nuclear proteins prepared from lactating guinea pig mammary glands on probes derived from the guinea pig α-lactalbumin gene promoter. These complexes were however also formed by proteins prepared from the liver and kidney of lactating guinea pig, and therefore the In Vitro approach was judged to be non-instructive in this instance. Two lines of transgenic mice were constructed by microinjection of a 3.5kb guinea pig α-lactalbumin genomic fragment, and both lines were found to express the transgene at levels commensurate with the levels seen in the guinea pig. Guinea pig α-lactalbumin was secreted into the milk of transgenic mice. The transgene mRNA was correctly initiated and processed, and was found to be expressed at high levels exclusively in the mammary glands, and the skin of the transgenic mice. Both transgenic and non-transgenic lactating mice were observed to express the endogenous mouse α-casein gene in the skin. Guinea pig α-lactalbumin protein was not detected in the skin of lactating transgenic mice. In Situ hybridisation studies revealed that the expression of the transgene in the mammary tissues of transgenic mice was restricted to the secretory acini of the mammary glands, which is the expected site for milk protein production. In the skin, the expression of guinea pig α-lactalbumin mRNA was found to be confined to the sebaceous glands, more specifically to the ducts formed between the sebaceous gland Itself and the hair follicle.

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