人類體中有一個新穎的基因為zbrk1，其蛋白約為58kDa，此蛋白的特徵是N端與中間各帶有KRAB (Krüppel-associated box) 區域，以及八個鋅手指區域。ZBRK1另一個名稱是ZNF350 (zinc finger protein350，鋅手指蛋白350)，並且其功能上會與BRCA1作用影響下游基因已知透過特定之ZBRK1的特定DNA序列（GGGxxxCAGxxxTTT） ，ZBRK1能與BRCA1和CtIP結合成為複合體，進而結合在ANG1上的啟動子，抑制該基因表現，而能扮演抑制腫瘤生成的功能。然而，人類ZBRK1基因雖已在已被報導，但老鼠體內相對應的同源蛋白基因卻至今未知，因此在本研究嘗試利用蛋白質體研究方法將老鼠體內可能的同源蛋白找出，以利生醫領域學者得以依此進行後續研究並進一步釐清其功能。這裡所利用的蛋白質體策略是由下而上的方式去尋找此目標蛋白。我們使用反相管柱分離蛋白萃取液，並透過ZBRK1抗體鑑定出老鼠體內可能的ZBRK1同源蛋白所存在的蛋白分離液，並將此分離出的樣品使用蛋白酶酵素切割分解經由質譜儀分析進行蛋白質身份鑑定，最後使用生物資訊工具軟體，在老鼠蛋白質體資料庫中鑑定出可能的候選蛋白。然而低量的蛋白以及複雜生物樣品是鑑定老鼠體內ZBRK1的瓶頸，故在此我們著重於萃取條件和分析純化平台做為成果的呈現。

ZBRK1 gene, encodes a 58 kDa protein with an N-terminal KRAB (Krüppel-associated box) domain and eight central zinc fingers. The ZBRK1, also called ZNF350, is known to interact with BRCA1. Thus, BRCA1 can regulate it downstream targets through the consensus ZBRK1-binding motif, GGGxxxCAGxxxTTT. The previous studies discovered that the removal of BRCA1/CtIP/ZBRK1 repressor complex on ANG1 promoter lead to promoting tumor growth. Although human ZBRK1 is known, the mouse ZBRK1 has not been identified as yet. To identify mouse ZBRK1 for further studies, we try to find out human ZBRK1-homologue protein in mouse by a bottom-up proteomics approach. Combining with the identification of western blot by ZBRK1 antibody, we used reverse phase butyl column to enrich mouse homologous proteins. The result of western blot showed that ZBRK1 can be fractionated and concentrated in two fractions among a total of thirty fractions collected through a gradient elution by acetonitrile. These two mZBRK1 fractions were digested by trypsin and analyzed by LC/MS/MS. Based on protein identification results by using in-house zinc finger protein database we were able to deduce some protein sequences were the potential ZBRK1 homologue in mouse. However, low abundant mZBRK1 expression became a limiting factor in the identification of the mouse ZBRK1. Therefore, the extraction condition and the plateform of protein purification are the major points in this thesis.

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