论文部分内容阅读
目的克隆TRAF3IP3基因,明确TRAF3IP3在部分人类组织中的表达谱并鉴定其表达产物的亚细胞定位。方法采用实时定量PCR检测TRAF3IP3基因在人体各组织中的表达情况;克隆TRAF3IP3基因开放读码框区(ORF)并构建真核亚细胞定位质粒。转染人胚肾HEK 293细胞,激光共聚焦显微镜观察融合蛋白的亚细胞定位。结果实时定量PCR证实TRAF3IP3基因在被检测的11种组织中均有表达,其中在淋巴结、脾、骨髓、胃和睾丸中的表达水平较高;成功克隆了TRAF3IP3基因ORF区并构建成为荧光定位质粒,激光共聚焦显微镜观察证实TRAF3IP3蛋白主要定位于核膜,在核周呈颗粒状分布。结论 TRAF3IP3基因在淋巴细胞富集的器官呈高表达,可能参与免疫系统的发育、成熟及调控,其表达产物在细胞内主要定位于核膜及核周胞浆。
Objective To clone the TRAF3IP3 gene, clarify the expression profile of TRAF3IP3 in some human tissues and identify the subcellular localization of its expressed product. Methods Real-time quantitative PCR was used to detect the expression of TRAF3IP3 in human tissues. The open reading frame (ORF) of TRAF3IP3 gene was cloned and the eukaryotic subcellular localization plasmid was constructed. Human embryonic kidney HEK 293 cells were transfected and confocal laser scanning microscopy was used to observe the subcellular localization of the fusion protein. Results Real-time quantitative PCR confirmed that the TRAF3IP3 gene was expressed in all the 11 tissues tested. The expression level of TRAF3IP3 gene was higher in lymph nodes, spleen, bone marrow, stomach and testes. The ORF region of TRAF3IP3 gene was cloned successfully and constructed into fluorescently oriented plasmid Confocal laser scanning microscopy confirmed TRAF3IP3 protein located mainly in the nuclear membrane, in the nucleus was granular distribution. Conclusion The TRAF3IP3 gene is highly expressed in lymphocyte-enriched organs and may be involved in the development, maturation and regulation of the immune system. The expression of TRAF3IP3 mainly locates in the nuclear membrane and perinuclear cytoplasm.