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热适应差异表达基因消减文库的筛选
PCR-select differential screening of a subtracted cDNA library of heat-adapted rat liver

作  者: ; ; ; ;

机构地区: 中国人民解放军第一军医大学热带军队卫生学系

出  处: 《中国病理生理杂志》 2005年第10期2050-2053,共4页

摘  要: 目的:研究热适应大鼠的肝细胞基因差异表达,探讨热适应的分子机制。方法:构建热适应差异表 达基因消减文库,扩增并克隆T/A载体形成重组质粒,转化感受态细胞后分离获得目的基因,采用PCR-select dif ferential screening技术筛选差异表达基因,测序后所得序列与GenBank中的已知序列进行比对,确定其可能的功能。 结果:确认8个上调表达基因,与GenBank中已知序列进行比较,证实其中有3种已知基因,5个新序列表达标签 (EST)。结论:相关基因表达水平上调可能参与生物体热适应的信号转导通路,而p53可能是热适应分子环路的又一 节点。 AIM: To reveal the molecular mechanisms of heat adaptation by study the change of gene expression in rat liver when the rats exposed to hyperthennia stress and heat adaptation. METHODS: Differentially expressed genes in rat liver in association with heat adaptation were identified using the suppression subtractive hybridization (SSH) technique to prepare subtracted eDNA libraries in heat adaptation. The PCR- amplified eDNA fragments generated by SSH were then ligated into the plasmid pGEM - T (Promega). PCR - select differential screening technique was used to filtrate differentially expressed genes. Each of these dif- ferentially expressed genes was sequenced and was compared to known sequences in the GenBank database. RESULTS: eight upregulating expressed genes segments were isolated from the up - regulating differentially expressed library, three of which were known gene such as p53 and five genes were new expressed sequence tags. CONCLUSION: Up- regulating expressed genes such as 1353 maybe participate in the signal transduction pathway of heat adaptation.

关 键 词: 热适应 基因文库 数据库 核酸

领  域: [生物学]

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