【摘要】 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。更多还原

【Abstract】 ObjectiveTo construct and identify the transgenic mice with extrinsic fat-1 gene.MethodsFat-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 G0 generation of fat-1 transgenic mice.ResultsRecombinant plasmid pEF-fat-1 was successfully constructed and four G0 mice with integrated fat-1 gene were identified via PCR and Southern blot.ConclusionFat-1 gene can be integrated into mouse to obtain the transgenic mouse that provides an animal model for the study of fat-1 gene.更多还原