title: The high-level expression of human tissue plasminogen activator in the milk of transgenic mice with hybrid gene locus strategy.
abstract: Transgene expression for the mammary gland bioreactor aimed at producing recombinant proteins requires optimized expression vector construction. Previously we presented a hybrid gene locus strategy, which was originally tested with human lactoferrin (hLF) as target transgene, and an extremely high-level expression of rhLF ever been achieved as to 29.8 g/l in mice milk. Here to demonstrate the broad application of this strategy, another 38.4 kb mWAP-htPA hybrid gene locus was constructed, in which the 3-kb genomic coding sequence in the 24-kb mouse whey acidic protein (mWAP) gene locus was substituted by the 17.4-kb genomic coding sequence of human tissue plasminogen activator (htPA), exactly from the start codon to the end codon. Corresponding five transgenic mice lines were generated and the highest expression level of rhtPA in the milk attained as to 3.3 g/l. Our strategy will provide a universal way for the large-scale production of pharmaceutical proteins in the mammary gland of transgenic animals.
authors.affiliation: Cell Engineering Department, Beijing Institute of Biotechnology, Beijing 100071, China.
authors.forename: Yanrong
authors.initials: Y
authors.lastname: Zhou
authors.forename: Yanli
authors.initials: Y
authors.lastname: Lin
authors.forename: Xiaojie
authors.initials: X
authors.lastname: Wu
authors.forename: Fuyin
authors.initials: F
authors.lastname: Xiong
authors.forename: Yuemeng
authors.initials: Y
authors.lastname: Lv
authors.forename: Tao
authors.initials: T
authors.lastname: Zheng
authors.forename: Peitang
authors.initials: P
authors.lastname: Huang
authors.forename: Hongxing
authors.initials: H
authors.lastname: Chen
mesh_terms: D000818:Animals; D005260:Female; D005786:Gene Expression Regulation; D005818:Genetic Engineering / Q000379:methods; D006801:Humans; D008321:Mammary Glands, Animal / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic / Q000235:genetics*; D008892:Milk / Q000378:metabolism*; D008894:Milk Proteins / Q000235:genetics; D010959:Tissue Plasminogen Activator / Q000235:genetics* / Q000378:metabolism; D019076:Transgenes
chemical_list: D008894:Milk Proteins; C024471:whey acidic proteins; D010959:Tissue Plasminogen Activator