Abstract: Abstract：Objective Use escherichia coli B·galactasc gene LacZ as report gene，established Tyr03，Axl receptor tyrosine kinases transgenic mouse containing loxp sites，mating with different tissue specific Cre mice to obtain space．time expressions of transgenic mice，make a foundation for research Tyr03 and Axl in each organization functions．Methods Constructing Tyr03，Axl receptor tyrosine kinase plasmid containing loxp sites，sequencing correctly，through microscopic injection technology to insert the Geo-STOP—msAxl and Geo—STOP。msTyr03 gene containing CAG promoter upstream into BDFl fertilized eggs，Transplant to ICR receptor mouse，Stay after breeding， using double primers PCR to appraisal Fo and Fn generation mouse genotype，using RT·PCR，X—gal staining to observe LacZ genes’express in mice organization，using WB to detect the protein expression in transgenic mice and wild type．Result Adopt double primer PCR to appraisal new born mice，get modified positive transgenic mouse， use the RT．PCR conformed the exist of LacZ gene；the x—gal staining，observed blue precipitation in the brain， testicular and the thymus of Geo—STOP—msTyr03。observed blue precipitation in brain，heart，ovarian，kidney of Geo· STOP．msAxl，indirectly illustrate the expression part of aim gene；For identification of double primers positive Geo· STOP．msTyr03。Geo—STOP．msAxl transgenic mice and wild type mice，we use the WB to validate the Axl and Tyr03 gene expressing no difference，and explain the exist of STOP sequence，so the Axl and Tyr03’8 expression is restrained．Conclusions Successful got Geo—STOP．msAxl and Geo·STOP—msTyr03 tyrosine receptor transgenic moUSe．

Key words: Tyr03, Axl receptor tyrosine, Loxp site, X—gal staining

摘要： 目的采用大肠杆菌B-半乳糖酶基因LaeZ作为报告基因。建立Tyr03、Axl受体酪氨酸激酶含有loxp位点的 转基因小鼠。与不同组织特异性Cre小鼠杂交，获得时空性表达Tyr03、Axl的转基因小鼠，为研究其在各个组织中 的功能奠定基础。方法构建含有Ioxp位点的Tyr03、Axl受体酪氨酸激酶质粒，测序正确后，通过显微注射法将插 入CAG启动子下游的Geo·STOP—msAxl及Geo—STOP—msTyr03基因的转基因载体注射人BDFI供体鼠受精卵，移植 受精卵至ICR受体鼠，待产仔后，通过双引物PCR鉴定F。以及Fn代小鼠的基因型。RT-PCR，X．gal染色观察小鼠 组织中LacZ基因，运用WB(Western Blot)检测转基因小鼠和野生型小鼠体内目的蛋白表达的差异。结果获得了 含有目的基因的转基因阳性小鼠；通过RT-PCR对阳性小鼠证明LacZ基因的存在；对存在LacZ基因的小鼠进行x． gal染色，在Geo-STOP-msTyr03的脑、睾丸、胸腺中观察到蓝色沉淀，在Geo-STOP-msAxl的脑、心、肾中观察到蓝色 沉淀，间接说明目的基因能表达的组织和器官；对双阳性的Geo-STOP．msTyr03、Geo-STOP—msAxl转基因小鼠和野生 型小鼠，WB检验证实Tym3、Axl基因的表达没有差异，说明在加入stop序列之后，Tym3基因确实存在，而且表达 受到抑制。结论成功建立Geo—STOP—msAxI及Geo-STOP-msTyr03酪氨酸受体转基因小鼠。

关键词: Tyr03、Axl受体酪氨酸激酶, loxp位点, X．gal染色