人ApoE转基因小鼠与TGE_（7－2）　TGE_（7－3）转基因鼠系的建立

Transgenic Mice of Human ApoE_(7-2) and ApoE_(7-3)

用含小鼠金属硫蛋白（ＭＴ－１）基因启动子与突变的人ＡｐｏＥ７基因的６．０ＫｂＤＮＡ片段作为目的基因制备转基因小鼠，利用显微注射法将目的基因导入４４１枚受精卵，移植于２０只假孕鼠，其中１５只受孕，共产仔鼠８０只，经ａ－３２Ｐ斑点杂交与Ｓｏｕｔｈｅｒｎ杂交法鉴定出两只整合有人ＡｐｏＥ基因的转基因雌鼠，其拷贝数分别为２和５。将两只首建鼠（雌性Ｆｏ代）与正常雄鼠交配，建立Ｆ１代转基因鼠系ＴＧＥ７－２和ＴＧＥ７－３，为进一步从整体上研究ＡｐｏＥ在脂质代谢中的作用以及ＡｐｏＥ与动脉粥样硬化和Ａｌｚｈｅｉｍｅｒ’ｓ病的关系创造条件。

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.