心脏形成关键基因-TBX5在大鼠胚胎心脏中的克隆及表达研究(英文)  

Cloning of TBX5, a Key Gene During Heart Formation and Its Expression in Rat Embryonic Heart

作　　者：宫立国[1] 邱广蓉[1] 辛娜[1] 徐小延[1] 孙开来[1] 

机构地区：[1]中国医科大学遗传教研室,沈阳110001

出　　处：《Acta Genetica Sinica》2006年第3期199-205,共7页

基　　金：This work was supported by the National Natural Science Foundation of China (No. 30070411, 30200305 and 30400485) the Education Department Science Funds of Liaoning Province (No. 202013133, 2004C045).

摘　　要：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。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.

关 键 词：TBX5基因 克隆 表达 心脏发育 大鼠 

分 类 号：Q78[生物学—分子生物学]