应用抑制性消减杂交技术研究大鼠肝脏热应激过程中基因的差异表达
Zhongguo ying yong sheng li xue za zhi = Zhongguo yingyong shenglixue zazhi = Chinese journal of applied physiology（2003）
Abstract
目的:应用抑制性消减杂交技术,分离热应激过程中出现的差异表达基因,筛选与热应激相关的功能基因片断,探索热应激的分子机理.方法:实验分两组:热应激组和常温对照组,分别提取肝组织mRNA后反转录cDNA.内切酶酶切、接头连接后,以热应激组cDNA为tester,对照组cDNA为driver进行SSH.产物克隆T/A载体构建重组质粒转化感受态细胞后进行分离筛选.结果:mRNA质优量足,反转录cDNA并成功酶切为500 bp左右片断.连接效率满意.高效杂交后成功构建消减文库,并初步从中分离获得180个片段.结论:应用SSH技术构建热应激差异表达基因消减文库为筛选热应激相关基因奠定了基础,对探索支配热应激反应的功能基因有积极意义.
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Key words
RNA methylation,Gene Expression Regulation,mRNA modification
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