神经特异性过表达Omi转基因鼠的建立

目的建立神经特异性过表达Omi的小鼠模型。方法将神经元特异性烯醇化酶启动子(neu-ron-specific enolase,NSE)连接到Omi cDNA上游获得了pSPORT6.0-NSE-Omi重组质粒,并转染神经细胞验证过表达情况。利用EcoRV和PvuI双酶切获得4.3Kb的转基因片段,利用显微注射技术将基因片段注射到800枚受精卵中,其中挑选500枚移植至假孕鼠的输卵管中发育。获得的子代小鼠利用特异性引物PCR验证基因整合情况。结果显微注射后共获得子代小鼠114只,基因型鉴定有11只小鼠基因上有转基因整合基因,整合率为10%。结论通过重组质粒及显微注射技术可以获得神经特异性过表达Omi转基因鼠模型。

Establishment of a transgenic mice model with neuron-specific over-expression of Omi

Objective To establish a transgenic mice model which specific over-expressing of human Omi in nervous system.Methods The pSPORT6.0-NSE-Omi plasmid containing human Omi downstream of NSE(please tell us the whole name of NSE) promoter,which expresses specifically in nervous system,was obtained.The 4.3 Kb EcoRV/PvuI fragment from the recombinant plasmid was introduced into 800 zygotes by microinjection and 500 injected fertilized eggs were transferred into pseudopregnant recipients.Potential transgenic founder mice were identified by PCR with specific primers.Results One hundred and fourteen offsprings were obtained after transplantation.Eleven transgenic mice were identified by genotyping.Integration efficiency is 10%.Conclusions Transgenic mice with specific over-expressing Omi in nervous system are established.