【摘要】 背景:HOX基因家族是一个高度保守的转录因子家族,该家族在胚胎发育阶段基本体轴和次级体轴的形成中起作用,同时在中枢神经系统、中轴骨、胃肠和泌尿生殖系及外生殖器和肢体的发育中起重要作用。目的:观察孕14周人胚胎足部发育过程中与HOXD13相互作用的蛋白质。方法:取胎儿足部组织,应用蛋白提取试剂盒提取蛋白,加入6μgHOXD13抗体进行免疫沉淀实验获取HOXD13相互作用的蛋白质,以加入非特异的IgG作为阴性对照。考马斯亮蓝染色后观察沉淀下来蛋白质形成的条带,切取需要条带送质谱分析。结果与结论:利用HOXD13抗体在孕14周人胚足部组织蛋白质中获取了一个可能与HOXD13相互作用的蛋白质,相对分子质量约为60000,经质谱鉴定后为锌指蛋白548。在孕14周人胚足部发育过程中,HOXD13可能和锌指蛋白548结合形成蛋白复合体而发挥转录调控作用。更多还原

【Abstract】 BACKGROUND: The HOX gene family is a highly conserved transcription factor family, which affects the formation of basic axis and secondary axis during embryonic development, at the same time, it plays a pivotal role in the development of the central nervous system, axial skeleton, stomach intestine, urogenital system and external genitalia. OBJECTIVE: To investigate the interactional protein of HOXD13 in human embryo foot development at 14 weeks pregnancy. METHODS: Fetal foot tissues were harvested and protein extraction was performed using protein extraction kit, 6 μg HOXD13 antibody was added for immunoprecipitation experiment, which aims to obtain HOXD13 interactional proteins, while the non-specific IgG served as a negative control. Subsequent to Bradford staining, the sediment protein bands were cut for mass spectrometry. RESULTS AND CONCLUSION: We obtained an interactional protein of HOXD13 in the human embryo foot at 14 weeks pregnancy, at the relative molecular mass of approximately 60 000. The protein was identified as zinc finger protein 548 using mass spectrum. HOXD13 may play a very important role in transcriptional control by HOXD13-zinc finger protein 548 protein complex in the human embryo foot at 14 weeks pregnancy.更多还原