High-level expressing YAC vector for transgenic animal bioreactors.
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 alpha-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 alpha-lactalbumin gene was replaced with the hGH gene, tissue specificity of hGH mRNA was the same as that of the endogenous rat alpha-lactalbumin gene. Thus, the 210-kb human alpha-lactalbumin YAC is a useful vector for high-level expression of foreign genes in the milk of transgenic animals.
Fujiwara
Y
Y
YS New Technology Institute, Inc., Ishibashi-machi, Shimotsuga-gun, Tochigi, Japan.
Miwa
M
M
Takahashi
R
R
Kodaira
K
K
Hirabayashi
M
M
Suzuki
T
T
Ueda
M
M
D000818:Animals; D030801:Animals, Genetically Modified; D019149:Bioreactors; D018244:Chromosomes, Artificial, Yeast; D017931:DNA Primers; D005260:Female; D005821:Genetic Techniques; D005822:Genetic Vectors; D019382:Human Growth Hormone / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D007768:Lactalbumin / Q000235:genetics*; D008297:Male; D008892:Milk / Q000737:chemistry; D009928:Organ Specificity; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis; D014158:Transcription, Genetic
D017931:DNA Primers; D012333:RNA, Messenger; D011994:Recombinant Proteins; D019382:Human Growth Hormone; D007768:Lactalbumin