[Cloning of human G-CSF genomic gene and its expression in transgenic mice mammary gland].
Lu Yf, C Tian, Deng Jx, Xiao Cz, Ma Qj
Abstract
Lu Yf, C Tian, Deng Jx, Xiao Cz, Ma Qj
Abstract
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.
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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.
Key concepts: Biology, genomic DNA, Molecular biology, Gene, Transgene, Genetically modified mouse, Cloning (programming), Gene expression