小鼠Nucleophosmin基因的克隆及基因组结构分析  

Cloning and Structural Analysis of Mouse Genomic Nucleophosmin Gene

作　　者：项佑贵[1,2] 陆顺元[1] 顾鸣敏[1] 王升跃 任双喜 傅刚[3] 王铸钢[1,2] 

机构地区：[1]上海第二医科大学医学遗传学教研室 [2]上海南方模式生物研究中心,上海201203 [3]国家人类基因组南方研究中心

出　　处：《Acta Genetica Sinica》2005年第6期641-649,共9页

基　　金：国家杰出青年科学基金项目(编号:39925023);国家高技术研究发展计划项目(编号:2001AA216081);上海市科技发展基金项目(编号:99JC14029;99XD14005)资助~~

摘　　要：Nucleophosmin(NPM)是主要的核仁磷酸化蛋白,在一些退行性大细胞淋巴瘤、骨髓增生异常综合征、急性髓性白血病等病人中可见累及NPM基因的染色体易位。建立NPM基因剔除的小鼠模型对于研究NPM基因的生物学功能及其在淋巴瘤及白血病发病中的确切作用有非常重要的意义。为获得包含NPM基因组全长序列的噬菌体克隆,以小鼠NPMcDNA为探针对129S1系小鼠λ噬菌体基因组文库进行筛选,经3轮筛选鉴定出一个包含小鼠NPM基因组全序列的噬菌体克隆。用鸟枪法对这个λ噬菌体克隆全长15.3kb进行了测序,序列经BLAST分析显示,该克隆所包含的129S1系小鼠NPM基因组序列与GenBank中C57BL/6系小鼠序列约99.8%相同。根据测序获得的基因组序列,应用生物信息学方法对小鼠NPM的基因组结构及NPM基因5′启动子区可能的转录因子结合位点等进行了分析。Nucleophosmin (NPM) is an abundant nucleolar phosphoprotein. NPM gene involved chromosomal translocations were found in the patients with anaplastic large cell lymphomas(ALCL),myelodysplastic syndrome (MDS),acute myeloid leukemia (AML)and acute promyelocytic leukemia (APL).To generate NPM gene knockout mice and study its biological function in vivo ,we screened the λ phage genomic library derived from 129S1 mice with mouse NPM cDNA probe.A positive phage clone which contained the full length NPM genomic DNA was obtained and the insert of 15 3 kb genomic DNA in this clone was sequenced with shotgun method.BLAST analysis showed that the sequence of insert are 99 8% identity to that of NPM gene of C57BL/6 mouse strain.Based on the sequence,bioinformatics analysis on genomic structure of NPM and the transcription factor binding sites in the NPM 5′ flanking region were performed.

关 键 词：小鼠 Nucleophosmin基因 基因克隆 基因组结构 核仁磷酸化蛋白 Λ噬菌体 

分 类 号：Q78[生物学—分子生物学]