Transgenic Mice of Human ApoE_(7-2) and ApoE_(7-3)
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KeyWord:Apolipoprotein E Transgenic mice Mouse line,
Jiang Tian,et al
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      iven the stronbv association of Apolipoprotein E (ApoE )wiih Athe-rosclerosis (AS) and Alzheimer's Disease (AD), this project aims ofsetting upthe transgenicmice mice a mutant human ApoE gene. Thisspecial animal model would be great benefit to understandinato the cffect ofabnormal ApoE gene expression on lipids metabolism and its roles in thepalhogenesis of AS and AD.A 6.0kb DNA fragment from the recombinant piasmid PME7 containingthe mouse MT-I promoter and the human ApoE7 (Glu 244, 245-Lys 244,245) genomic DNA was recovered from low melting agarose electophoresisand purified by CsCI ultracentr ifugation.Then it was microinjected intothe male pronucleus of the fertilized eggs of KM mice. 441 eggs survivedfroni microinjection were transplanted into the oviduct of 20 pseudopregn-ant feiiiaie, 15 pseudopregnant nivice got pregnant and gave birth to 80pups. Genomic DNA were extracted from the tails of the pups Dot Blothybrid ization was first used for screening. Then Southern Blot hybridi-zation was used fordetermination of the integrationof h-ApoE, gene. A2. Okb DNA fragment including the forth exon of human ApoE7 genomicDNA was used as the probe. 2 mice wiare identified of the intergrationof h-ApoE7 gene with 2 and 5 copies respectively. The 2 founder femalstransgenic mice ( Fo generation ) were bred to ordinary KM male mice toset up Fi generation. One mouse aborted on the 17th day of pregnancy.while another one had successfully given birth to 10 pups which are nowready for identification.
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