The Construction Of The Vector And The Establishment Of Lines Of Transfected Goat Mammary Epithelial Cells In Vitro For Mammary Specific Expression Of Human Lactoferrin

Human lactoferrin (hLTF), also called lactotransferrin, a component of mammalian milk, is an iron-binding protein in host defense against infection and severve inflammation, regulation of the cell's development and ferrum metabolism, and so on.Therefore the research of producing recombinant hLTF(hLTF) by transgenic animal has many values for medical therapy and human health.Somatic cell cloning technology is successfully used for producing transgenic animals. Transfection of the heterogenous DNA into animal somatic cells, including adult animal's somatic cell and fetal blast cell, then transfer of this donor cell into an enucleated oocyte to produce reconstituted embryos are capable, upon transfer to suitable hosts, of developing into viable offspring.Under the base of the successful technique of somatic nuclear transfer and mammary gland bioreactor, this thesis systematically study on constructing mammary gland-specific expressional vector, establishing goat mammary epithelial cell lines in vitro, transferring foreign genes into cultured cells and establishing transfected cell clonies, as the following two steps.1 Construction of the mammary gland-specific expressional vectorFor the purpose of constructing the vector afLF, ligation the cDNA (2320bp) of human lactoferrin gene into 5' and 3' flank regulatory sequence of aS1-casein gene, and the Neo'(700bp) gene and the sequence of enhancer which had harbored in our laboratory is required. The human lactoferrin cDNA (hLTF) was amplified from the templet of human...