[Cloning of human G-CSF genomic gene and its expression in transgenic mice mammary gland].
Human genomic DNA was used as template of PCR. 1.5 kb G-CSF genomic DNA was obtained using PCR amplification method. Sequence analysis showed that genomic DNA sequence of human G-CSF was correct. The vector of mammary gland expression was constructed and contained whey acid protein (WAP) 5' control region directed human G-CSF genomic DNA. In order to produce transgenic mice, 1200 fertilized eggs were microninjected using WAP-G-CSF fragment. Two male transgenic mice were obtained and identified using PCR method and Southern analysis. Integration rate of human G-CSF gene was 2.37% in mice. Foreign gene could also be identified in F1 and F2 transgenic mice. Expression levels of human G-CSF in transgenic mouse milk were 120-250 ng/ml.
Lu
Y F
YF
Institute of Biotechnique Academy of Military Medicine, Beijing.
Tian
C
C
Deng
J X
JX
Xiao
C Z
CZ
Ma
Q J
QJ
D000818:Animals; D015139:Blotting, Southern; D003001:Cloning, Molecular; D005260:Female; D015870:Gene Expression; D016179:Granulocyte Colony-Stimulating Factor / Q000235:genetics*; D006801:Humans; D008297:Male; D008321:Mammary Glands, Animal / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D016133:Polymerase Chain Reaction
D016179:Granulocyte Colony-Stimulating Factor