CAJ | 학술논문

为了研究心脏形成关键基因-TBX5的表达情况,选择Wistar大鼠作为从分子水平研究心脏发育的实验动物并从胎鼠心脏中克隆了TBX5 cDNA全长(GenBank登录号:AY859491),其cDNA及氨基酸序列不同于GenBank预测序列;随后应用RT-PCR及Northern blot分析TBX5基因在大鼠各组织中的表达情况,发现TBX5为单一转录本,在大鼠各组织均有表达,心脏中表达最强;构建荧光表达载体,转染大鼠肝癌细胞,观察TBX5亚细胞定位,结果证实TBX5基因在细胞核中表达;最后构建原核表达载体,IPTG诱导表达获得了GST-TBX5融合蛋白.以上工作为进一步鉴定心脏发育中TBX5相关转录因子及研究相互间的调控机制奠定了基础.
위료연구심장형성관건기인-TBX5적표체정황,선택Wistar대서작위종분자수평연구심장발육적실험동물병종태서심장중극륭료TBX5 cDNA전장(GenBank등록호:AY859491),기cDNA급안기산서렬불동우GenBank예측서렬;수후응용RT-PCR급Northern blot분석TBX5기인재대서각조직중적표체정황,발현TBX5위단일전록본,재대서각조직균유표체,심장중표체최강;구건형광표체재체,전염대서간암세포,관찰TBX5아세포정위,결과증실TBX5기인재세포핵중표체;최후구건원핵표체재체,IPTG유도표체획득료GST-TBX5융합단백.이상공작위진일보감정심장발육중TBX5상관전록인자급연구상호간적조공궤제전정료기출.
To study the expression of TBX5, a key gene during heart formation, full-length TBX5 cDNA was cloned from Wistar rat embryonic heart (GenBank Accession No. AY859491). The cDNA and predicted amino acid sequences were different from those previously reported in GenBank. The expression profile of TBX5 in rat tissues was studied by RT-PCR and Northern blot. TBX5 was expressed in many rat tissues as a single transcript, and the highest level of expression was found in the heart. For subcellular localization, TBX5 was expressed as an EGFP fusion protein in rat hepatocarcinoma cells. Results showed that TBX5 was nuclear.In addition, TBX5-GST fusion protein was obtained by prokaryotic expression. These findings provide a good basis for further identification of TBX5-related transcription factors and protein-protein interaction studies among each other.