title: In vitro evaluation of a mammary gland specific expression vector encoding recombinant human lysozyme for development of transgenic dairy goat embryos.
abstract: A vector expressing human lysozyme (pBC1-hLYZ-GFP-Neo) was evaluated for gene and protein expression following liposome-mediated transformation of C-127 mouse mammary cancer cells. Cultures of G418-resistant clones were harvested 24-72 h after induction with prolactin, insulin and hydrocortisone. Target gene expression was analyzed by RT-PCR and Western blot and recombinant human lysozyme (rhLYZ) bacteriostatic activity was also evaluated. The hLYZ gene was correctly transcribed and translated in C-127 cells and hLYZ inhibited gram-positive bacterial growth, indicating the potential of this expression vector for development of a mammary gland bioreactor in goats. Guanzhong dairy goat skin fibroblasts transfected with pBC1-hLYZ-GFP-Neo were used to construct a goat embryo transgenically expressing rhLYZ by somatic nuclear transplantation with a blastocyst rate of 9.0 ± 2.8 %. These data establish the basis for cultivation of mastitis-resistant hLYZ transgenic goats.
authors.affiliation: Anhui Provincial Laboratory of Local Livestock and Poultry Genetical Resource Conservation and Breeding, College of Animal Science and Technology, Anhui Agricultural University, Hefei, 230036, Anhui, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Gui
authors.forename: Meiling
authors.initials: M
authors.lastname: Zhang
authors.forename: Jianwen
authors.initials: J
authors.lastname: Chen
authors.forename: Yuanliang
authors.initials: Y
authors.lastname: Zhang
authors.forename: Naru
authors.initials: N
authors.lastname: Zhou
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhang
authors.forename: Jia
authors.initials: J
authors.lastname: Tao
authors.forename: Liucai
authors.initials: L
authors.lastname: Sui
authors.forename: Yunsheng
authors.initials: Y
authors.lastname: Li
authors.forename: Ya
authors.initials: Y
authors.lastname: Liu
authors.forename: Xiaorong
authors.initials: X
authors.lastname: Zhang
authors.forename: Yunhai
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D019149:Bioreactors; D045744:Cell Line, Tumor; D004622:Embryo, Mammalian; D005260:Female; D005822:Genetic Vectors / Q000235:genetics*; D006041:Goats / Q000196:embryology / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D008297:Male; D008321:Mammary Glands, Animal / Q000166:cytology / Q000201:enzymology / Q000502:physiology*; D051379:Mice; D009113:Muramidase / Q000096:biosynthesis* / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D009113:Muramidase