带FLAG标签小鼠HuR真核表达载体的构建及其生物学功能的鉴定
带FLAG标签小鼠HuR真核表达载体的构建及其生物学功能的鉴定
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带FLAG标签小鼠HuR真核表达载体的构建及其生物学功能的鉴定
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Jie fang jun yi xue za zhi, 2011, Vol.36 (4), p.352-355
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Chinese
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目的构建人抗原R（HuR）的真核表达载体,观察其在NIH3T3细胞中的表达并初步分析其生物学功能。方法提取NIH3T3细胞总RNA并反转录为cDNA,以cDNA为模板PCR扩增得到小鼠HuR编码序列,酶切后克隆至pcDNA3.1-FLAG载体;重组载体经PCR、酶切、测序鉴定正确后瞬时转染NIH3T3细胞,采用Western blotting分析HuR在细胞中的表达,并采用Real-time PCR检测HuR过表达对DUSP1mRNA水平的影响。结果成功构建了重组质粒pcDNA3.1-HuR-FLAG,Western blotting显示该质粒能在NIH3T3细胞中高效表达,Real-time PCR结果表明HuR过表达可提高细胞内DUSP1基因的mRNA水平。结论成功构建了带FLAG标签的...
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带FLAG标签小鼠HuR真核表达载体的构建及其生物学功能的鉴定
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TN_cdi_chongqing_backfile_37666704
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https://devfeature-collection.sl.nsw.gov.au/record/TN_cdi_chongqing_backfile_37666704
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0577-7402
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