[Cloning and structural analysis of mouse genomic nucleophosmin gene].

You-Gui Xiang, Shun-Yuan Lu, Ming-Min Gu, Sheng-Yue Wang, Shuang-Xi Ren, Gang Fu, Zhu-Gang Wang
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Abstract

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 lambda 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.

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小鼠基因组核磷蛋白基因的克隆与结构分析
核磷蛋白(NPM)是一种丰富的核仁磷酸化蛋白。NPM基因参与染色体易位存在于间变性大细胞淋巴瘤(ALCL)、骨髓增生异常综合征(MDS)、急性髓系白血病(AML)和急性早幼粒细胞白血病(APL)患者中。为了构建NPM基因敲除小鼠并研究其在体内的生物学功能,我们使用小鼠NPM cDNA探针筛选了129S1小鼠的lambda噬菌体基因组文库。获得了含有NPM全长基因组DNA的噬菌体阳性克隆,用鸟枪法对该克隆中15.3 kb的基因组DNA片段进行了测序。BLAST分析结果表明,插入序列与C57BL/6小鼠品系NPM基因的同源性为99.8%。基于该序列,对NPM的基因组结构和NPM 5'侧区转录因子结合位点进行了生物信息学分析。
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