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High‐level expressing YAC vector for transgenic animal bioreactors
Author(s) -
Fujiwara Yoshihiro,
Miwa Masami,
Takahashi RiIchi,
Kodaira Kunihiko,
Hirabayashi Masumi,
Suzuki Takashige,
Ueda Masatsugu
Publication year - 1999
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/(sici)1098-2795(199904)52:4<414::aid-mrd10>3.0.co;2-s
Subject(s) - biology , transgene , gene , microinjection , microbiology and biotechnology , homologous recombination , genetically modified mouse , expression vector , recombinant dna , lactalbumin , position effect , genetics , biochemistry
The position effect is one major problem in the production of transgenic animals as mammary gland bioreactors. In the present study, we introduced the human growth hormone (hGH) gene into 210‐kb human α‐lactalbumin position‐independent YAC vectors using homologous recombination and produced transgenic rats via microinjection of YAC DNA into rat embryos. The efficiency of producing transgenic rats with the YAC vector DNA was the same as that using plasmid constructs. All analyzed transgenic rats had one copy of the transgene and produced milk containing a high level of hGH (0.25–8.9 mg/ml). In transgenic rats with the YAC vector in which the human α‐lactalbumin gene was replaced with the hGH gene, tissue specificity of hGH mRNA was the same as that of the endogenous rat α‐lactalbumin gene. Thus, the 210‐kb human α‐lactalbumin YAC is a useful vector for high‐level expression of foreign genes in the milk of transgenic animals. Mol. Reprod. Dev. 52:414–420, 1999. © 1999 Wiley‐Liss, Inc.