Gong Li-Guo, Qiu Guang-Rong, Xin Na, Xu Xiao-Yan, Sun Kai-Lai
Department of Medical Genetics, China Medical University, Shenyang 110001, China.
Yi Chuan Xue Bao. 2006 Mar;33(3):199-205. doi: 10.1016/S0379-4172(06)60040-1.
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.
为研究心脏形成过程中的关键基因TBX5的表达情况，从Wistar大鼠胚胎心脏中克隆了全长TBX5 cDNA（基因库登录号：AY859491）。该cDNA及预测的氨基酸序列与之前基因库中报道的不同。通过逆转录聚合酶链反应（RT-PCR）和Northern印迹法研究了TBX5在大鼠组织中的表达谱。TBX5在许多大鼠组织中以单一转录本形式表达，且在心脏中表达水平最高。为进行亚细胞定位，TBX5在大鼠肝癌细胞中作为绿色荧光蛋白（EGFP）融合蛋白表达。结果显示TBX5定位于细胞核。此外，通过原核表达获得了TBX5-谷胱甘肽S-转移酶（GST）融合蛋白。这些发现为进一步鉴定TBX5相关转录因子以及相互之间的蛋白质-蛋白质相互作用研究提供了良好基础。
