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人钙周期蛋白结合蛋白(hCacyBP)基因的克隆与表达
Cloning and Expression of Human Calcyclin Binding Protein (hCacyBP) Gene
【摘要】 筛选cDNA文库得到了人的钙周期蛋白结合蛋白基因 ,将此基因的全编码区克隆到原核表达载体pET2 8上 ,诱导目的蛋白质表达以后将重组蛋白质用亲和层析的方法进行纯化 ,得到了纯度很好的重组的目的蛋白质 ,以此作为抗原免疫动物 ,得到抗钙周期蛋白结合蛋白的特异多克隆抗体。Western印迹的结果表明 ,该基因在小鼠多种组织中广泛表达 ;免疫组化的结果表明 ,BT32 5细胞诱导分化后钙周期蛋白结合蛋白分布有变化 ,由分布于胞质中转向分布于胞核和核周胞质
【Abstract】 Human calcyclin binding protein (hCacyBP) gene was obtained by the screening of a human cDNA library. The full coding region of CacyBP was cloned into E.coli strain pET28, and then was expressed and purified through affinity chromatography. Rabbit anti-human CacyBP polyclonal antibody was obtained by immunizing rabbit with the purified human CacyBP. Western blots showed that it was expressed extensively in many tissues of mouse. The results of immunohistochemistrial staining showed that the location of CacyBP in BT325 cell line before and after differentiation changed from cytoplasm into nucleus and perinucleus cytoplasm.
【Key words】 calcyclin; calcyclin-binding protein (CacyBP); cloning; expression;
- 【文献出处】 生物化学与生物物理学报 ,Acta Biochimica Et Biophysica Sinica , 编辑部邮箱 ,2002年02期
- 【分类号】Q785;Q786
- 【被引频次】1
- 【下载频次】86