fat-1转基因小鼠的构建与鉴定

目的构建和鉴定携带有外源fat-1基因的转基因小鼠。方法将fat-1基因的cDNA与动物表达载体pEF—neo连接，构建pEF-fat-1重组质粒，酶切、测序鉴定正确后，以显微注射法把线性化重组质粒注射到小鼠受精卵的雄原核中，并将受精卵移植到受体鼠的输卵管中产出转基因小鼠，通过PCR、Southern blot杂交等方法确立阳性整合有目的基因的G0代小鼠。结果成功构建了pEF-fat-1重组质粒，将其显微注射到小鼠受精卵中，得到G0代小鼠，PCR、Southern blot杂交确立了4只整合有fat-1基因的首建鼠。结论fat-1基因可整合到小鼠体内，得到的转基因小鼠为研究fat-1基因的生物学功能提供了动物模型。

CONSTRUCTION AND IDENTIFICATION OF fat-1 TRANSGENIC MICE

Objective To construct and identify the transgenic mice with extrinsic fat-1 gene. Methods Fat-1 cDNA and animal expression vector pEF-neo were conjugated to construct the recombinant plasmid pEF-fat-1 which was digested by restrict enzyme and sequenced correctly. The obtained linear recombinant plasmid pEF-fat-1 was microinjected into the arsenoblasts of mouse zygote, which was implanted into the uterus to produce transgenic mice. PCR and Southern blot were performed to identify the positive Go generation of fat-1 transgenic mice. Results Recombinant plasmid pEF-fat-1 was successfully constructed and four Go mice with integrated fat-1 gene were identified via PCR and Southern blot. Conclusion Fat-1 gene can be integrated into mouse to obtain the transgenic mouse that provides an animal model for the study of fat-1 gene.