Fig. 3.

Generation of transgenic mice expressing mammalian target of Rap (mTOR) mutants. A: diagram of the mTOR transgene expression vector is shown. FLAG-tagged mTOR [Rap resistant (RR) or RR/kinase inactive (KI)] was under the control of the human skeletal muscle actin (HSA) promoter. The NruI-BstZ17I fragment was used for pronuclear injection. BamHI sites and the probe used for Southern analysis of the transgene integration are shown. B: representative Southern blot for genomic DNA isolated from RR and RR/KI mice tails is shown. C: transgene expression was examined by Western analysis of limb muscle homogenates, using anti-FLAG and anti-mTOR antibodies. Anti-tubulin blot served as a loading control.