文章摘要
王子露,苗登顺,季 吉.体外鉴定甲状旁腺激素相关蛋白与Bmi-1的相互作用[J].南京医科大学学报,2010,(12):1718~1721
体外鉴定甲状旁腺激素相关蛋白与Bmi-1的相互作用
Interaction between parathyroid hormone related protein with Bmi-1 in vitro
  
DOI:10.7655
中文关键词: 甲状旁腺激素相关蛋白  Bmi-1  GST pull-down
英文关键词: parathyroid hormone related protein  Bmi-1  GST pull-down
基金项目:中加合作项目(KY1010111091110254)
作者单位
王子露 南京医科大学口腔医学研究所,江苏 南京 210029 
苗登顺 南京医科大学口腔医学研究所,江苏 南京 210029 
季 吉 南京医科大学护理学院,江苏 南京 210029 
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中文摘要:
      目的: 探索并鉴定甲状旁腺激素相关蛋白(parathyroid hormone related protein,PTHrP)与Bmi-1的结合作用,为进一步研究PTHrP相关信号转导通路提供线索?方法:应用RT-PCR方法体外扩增鼠源性PTHrP基因,经测序后重组人原核表达载体pGEX-2TK,将构建正确的pGEX-2TK/mPTHrP原核表达载体转化E.coli BL21,IPTG诱导表达?表达产物经活性鉴定,通过体外GST pull-down技术和免疫印迹实验,检测PTHrP与Bmi-1蛋白的相互作用?结果:成功构建和表达了重组载体pGEX-2TK/mPTHrP;通过体外蛋白质结合实验证实了PTHrP与Bmi-1蛋白可以相互作用?结论:PTHrP与Bmi-1蛋白之间存在相互作用,此结果为研究PTHrP介导的下游信号转导通路的机制奠定了基础?
英文摘要:
      Objective: To identify the interaction between protein PTHrP and Bmi-1, and to further explore the underlying signaling pathway of PTHrP. Methods: The PTHrP gene was amplified by RT-PCR and inserted into corresponding sites of expression vector pGEX-2TK, and the fusion protein expressed in E.coli BL21 induced by IPTG. After identifying, the interactions between protein PTHrP and Bmi-1 were characterized by GST pull-down assay and Western Blot. Results:The new recombinant expression vector pGEX-2TK/mPTHrP was constructed and expressed successfully our. experiment confirmed the interactions between the protein PTHrP and Bmi-1. Conclusion:PTHrP specifically interacted with Bmi-1, which may modulate PTHrP downstream signaling pathway.
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