肝癌高表达新基因的克隆和编码蛋白质二级结构分析  

Clone of a novel liver cancer associated gene and analysis of the second structures of the predict protein

作　　者：王征旭[1] 胡桂芳[2] 王红阳[3] 吴孟超[3] 

机构地区：[1]兰州军区兰州总医院普外科,兰州730050 [2]兰州军区兰州总医院超声科,兰州730050 [3]第二军医大学东方肝胆外科医院

出　　处：《中华肝脏病杂志》2002年第1期25-27,共3页Chinese Journal of Hepatology

基　　金：国家自然科学基金(30000077;39825114);中国博士后科学资助金(中博基1999[10])

摘　　要：目的 研究和克隆新的肝癌相关基因，探索肝癌发生的分子机制。方法 采用ｍＲＮＡ差异显示技术和筛选胎盘ｃＤＮＡ文库，获得新基因全长ｃＤＮＡ；制备新基因的ＧＳＴ融台蛋白和多克隆抗体，进行ｗｅｓｔｅｒｎ杂交和免疫组织化学染色；利用计算机软件分析新基因的二级结构和功能预测。结果克隆了一个新的肝癌相关基因全长ｃＤＮＡ，ｗｓｔｅｒｎ杂交证实该基因全长能在２９３细胞中表达，免疫组织化学染色证实该基因编码蛋白定位于细胞浆，二级结构分析发现含２个ＳＨ３结台结构域和数个不同的蛋白激酶磷酸化位点。结论 获得一个新的肝癌相关基因全长ｃＤＮＡ。Objective To clone a novel liver cancer associated gene, and to explore the molecular basis of liver cancer genesis. Methods Using mRNA differential display polymerase chain reaction (DDPCR) and screening the human placenta cDNA library, we got a full-length cDNA of the gene. We prepared and purified the GST fusion protein and the special polyclonal antibody, engaged in the Western blot and immunohistochemical staining analysis, and analyzed the second structures and predicted the function of the protein by the computer soft. Results We have got a full-length cDNA of the liver cancer associated gene and identified that the full-length eDNA of the gene could be expressed in 293 eukaryocytes by Western blot assay. We localized the target protein in cytoplasm using the immunohistochemical staining methods, and found two SH3 binding domains and several protein kinase phosphorylation sites by analyzing the second structures. Conclusions We have got a novel full-length eDNA of human liver cancer associated gene.

关 键 词：肝细胞癌 基因克隆 基因表达 编码蛋白质 二级结构 DDPCR MRNA 

分 类 号：R735.7[医药卫生—肿瘤]