STAT3入核分子机制的初步研究

The Mechanism of Stat3 Nuclear Import

为了研究Stat3入核的分子机制 ,将SV40大T抗原的经典核定位序列NLS(nuclearlocalizationsequence)分别融合在Stat3 GFP分子和缺失突变体Dstat3 GFP的分子之间 ,构建Stat3 NLS GFP和Dstat3 NLS GFP融合分子。转染2 93T细胞 ,以NLS GFP为阳性对照 ,通过激光共聚焦显微镜的观察融合分子的亚细胞位置 ,未经白介素 6刺激的Stat3 NLS GFP和经白介素 6刺激的Stat3 GFP呈胞核分布 ,未经白介素 6刺激的Stat3 GFP和Dstat3 NLS GFP呈胞浆分布 .

In order to investigate the mechanism of stat3 nuclear import. positio ned a characterized NLS of the SV40 large T antigen into Stat3\|GFP ,Dstat3\|GF P respectively between the C\|terminus of Stat3 and the N\|terminus of GFP to cr eate Stat3\|NLS\|GFP and Dstat3\|NLS\|GFP. With NLS\|GFP as the positive contro l, Expression of the Stat3\|NLS\|GFP without IL\|6 stimulation and Stat3\|GFP wi th IL\|6 stimulation resulted in a predominantly nuclear localization in 293T ce ll. Expression of Stat3\|GFP and Dstat3\|NLS\|GFP without IL\|6 stimulation resu lted in predominantly cytoplasm localization in 293T cell. The results suggest that latent Stat3 is not anchored in the cytoplasm, and that nuclear localizatio n in response to IL\|6 is facilitated by gain of an NLS function.