title: Validation of a recombinant human bactericidal/permeability-increasing protein (hBPI) expression vector using murine mammary gland tumor cells and the early development of hBPI transgenic goat embryos.
abstract: Human bactericidal/permeability-increasing protein (hBPI) is the only antibacterial peptide which acts against both gram-negative bacteria and neutralizes endotoxins in human polymorphonuclear neutrophils; therefore, hBPI is of great value in clinical applications. In the study, we constructed a hBPI expression vector (pBC1-Loxp-Neo-Loxp-hBPI) containing the full-length hBPI coding sequence which could be specifically expressed in the mammary gland. To validate the function of the vector, in vitro cultured C127 (mouse mammary Carcinoma Cells) were transfected with the vector, and the transgenic cell clones were selected to express hBPI by hormone induction. The mRNA and protein expression of hBPI showed that the constructed vector was effective and suitable for future application in producing mammary gland bioreactor. Then, female and male goat fibroblasts were transfected with the vector, and two male and two female transgenic clonal cell lines were obtained. Using the transgenic cell lines as nuclear donors for somatic cell nuclear transfer, the reconstructed goat embryos produced from all four clones could develop to blastocysts in vitro. In conclusion, we constructed and validated an efficient mammary gland-specific hBPI expression vector, pBC1-Loxp-Neo-Loxp-hBPI, and transgenic hBPI goat embryos were successfully produced, laying foundations for future production of recombinant hBPI in goat mammary gland.
authors.affiliation: Anhui Provincial Laboratory of Animal Genetic Resources Protection and Breeding, College of Animal Science and Technology, Anhui Agricultural University, No. 130 of Changjiang West Road, Hefei, Anhui Province 230036, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Gui
authors.forename: Xing
authors.initials: X
authors.lastname: Liu
authors.forename: Jia
authors.initials: J
authors.lastname: Tao
authors.forename: Jianwen
authors.initials: J
authors.lastname: Chen
authors.forename: Yunsheng
authors.initials: Y
authors.lastname: Li
authors.forename: Meiling
authors.initials: M
authors.lastname: Zhang
authors.forename: Ronghua
authors.initials: R
authors.lastname: Wu
authors.forename: Yuanliang
authors.initials: Y
authors.lastname: Zhang
authors.forename: Kaisong
authors.initials: K
authors.lastname: Peng
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; D023181:Antimicrobial Cationic Peptides / Q000235:genetics* / Q000378:metabolism; D001798:Blood Proteins / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D019976:Cloning, Organism / Q000662:veterinary*; D004622:Embryo, Mammalian; D005260:Female; D005822:Genetic Vectors; D006041:Goats / Q000196:embryology / Q000235:genetics*; D006801:Humans; D008325:Mammary Neoplasms, Experimental / Q000235:genetics / Q000473:pathology*; D051379:Mice; D014162:Transfection; D019076:Transgenes; D054928:Validation Studies as Topic
chemical_list: D023181:Antimicrobial Cationic Peptides; D001798:Blood Proteins; C021623:bactericidal permeability increasing protein
keywords: C-127 cell line; Human bactericidal/permeability-increasing protein; Mammary gland bioreactor; Nuclear transfer; Transgene