条件性SV40 TAg转基因小鼠模型的建立

Generation of conditional simian virus 40 large T antigen gene transgenic mouse model

目的建立可调控的脑组织特异性表达SV40 TAg的转基因小鼠模型。方法构建由大鼠神经元特异性烯醇化酶基因(rat neuron-specific enolase,NSE)启动子驱动四环素基因表达调控系统的载体,从而控制SV40 TAg基因的表达;受精卵雄原核显微注射该线性化载体制备转基因小鼠;PCR方法鉴定首建鼠;RT-PCR方法检测小鼠组织中四环素反式激活子(rtTA)及SV40 TAg基因的表达情况。结果显微注射获得17个首建鼠,其中的6个首建鼠建系成功;1#、6#品系小鼠的脑组织中表达rtTA基因;强力霉素诱导后,可检测到6#品系小鼠脑组织中SV40TAg基因的特异性表达。结论可调控的脑组织表达SV40 TAg的转基因小鼠模型已经建立。

Objective To create a genetic mouse model with inducible expression of simian virus 40 large T antigen gene （ SV40 TAg） specifically in brain. Methods A single plasmid vector containing the regulatory element and TAg expression element of Tet-On expression system driven by rat neuron-specific enolase （NSE） promoter was constructed and micro-injected into the male pronuclei of fertilized ova from F1 hybrid mice （C57BL/6J × CBA）. The founder mice were identified by PCR analysis. The expression of rtTA mRNA and SV40 TAg in different tissues were examined by RT-PCR. Results Seventeen founder mice carrying both rtTA and TAg genes were identified by PCR in which six Tet-On-NSE-T genetic transgenic mouse lines were successfully established. Expression of rtTA mRNA was detected in the brains of mice from line 1^# and 6^#. SV40 TAg mRNA was expressed in the brain tissue from mouse line 6^# after intraperitoneal injection of Doxycycline. Conclusion A genetic mouse model with inducible expression of SV40 TAg in brain was successfully established.