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dc.contributor.authorWu, HTen_US
dc.contributor.authorLin, CSen_US
dc.contributor.authorHuang, MCen_US
dc.date.accessioned2014-12-08T15:41:32Z-
dc.date.available2014-12-08T15:41:32Z-
dc.date.issued2003en_US
dc.identifier.issn1031-3613en_US
dc.identifier.urihttp://hdl.handle.net/11536/28245-
dc.description.abstractThe 5'-regulative sequence and intron 1 of the goat beta-casein gene from -4044 to +2123 by was cloned and fused with the reporter gene of green fluorescent protein (GFP) to create a plasmid termed pGB562/GFP To detect GFP expression, pGB562/GFP was transfected in vitro via liposomes into the mammary epithelial cell line NMuMG. Cells could not express GFP unless the transfected NMuMG cells lined up to create functional alveoli. These functional cells were cultured with lactogenic hormones, including insulin, dexamethasone and prolactin, and were grown on a layer of the extracellular matrix Matrigel. Green fluorescent protein expression levels in NMuMG cells were 25-, 55- and 42-fold those in the control group at 24, 48, and 72 h after pGB562/GFP transfection respectively. In addition, pGB562/GFP was transfected ex vivo by electroporation into mammary gland fragments and cells were then cultured in vitro with a supplement of lactogenic hormones. Strong GFP expression localized in fragments of the mammary gland was observed 24 h after gene transfer. The novel strategy of ex vivo gene transfer into mammary tissue using GFP as a reporter gene to detect the function of a tissue-specific promoter is efficient and convenient. The data obtained herein reveal that the 5'-regulative sequence and intron 1 of the 6.2 kb goat beta-casein gene can enhance the efficiency of transgene expression. Thus, the GB562 sequence may act as a good promoter and effectively elevate the production of exogenous protein in mammary glands.en_US
dc.language.isoen_USen_US
dc.titleIn vitro and ex vivo green fluorescent protein expression in alveolar mammary epithelial cells and mammary glands driven by the distal 5 '-regulative sequence and intron 1 of the goat beta-casein geneen_US
dc.typeArticleen_US
dc.identifier.journalREPRODUCTION FERTILITY AND DEVELOPMENTen_US
dc.citation.volume15en_US
dc.citation.issue4en_US
dc.citation.spage231en_US
dc.citation.epage239en_US
dc.contributor.department生物科技學系zh_TW
dc.contributor.departmentDepartment of Biological Science and Technologyen_US
dc.identifier.wosnumberWOS:000184733300011-
dc.citation.woscount9-
Appears in Collections:Articles