Construction and expression of human thioredoxin eukaryotic expression vector
邹向晖, 邹宁, 周令望, 于维汉
Abstract
邹向晖, 邹宁, 周令望, 于维汉
Abstract
目的 构建人硫氧还蛋白基因的重组真核表达载体.方法人硫氧还蛋白的cDNA克隆至pGEM-T Easy载体,经DNA测序证明后,将其克隆至真核表达载体pShttle构建重组pShttle-hTRX,pShttle-hTRX转染Hela细胞系进行瞬时表达,通过Westem blot、免疫组化、活性测定、酶切、PCR扩增等方法证实构建载体的正确性.结果构建携带人硫氧还蛋白基因的真核表达载体pShttle-hTRX,转染HeLa细胞后可检测到hTRX的转录和表达.结论正确构建了携带人硫氧还蛋白基因的真核表达载体,本结果可用于探讨硫氧还蛋白的生物学作用。
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目的 构建人硫氧还蛋白基因的重组真核表达载体.方法人硫氧还蛋白的cDNA克隆至pGEM-T Easy载体,经DNA测序证明后,将其克隆至真核表达载体pShttle构建重组pShttle-hTRX,pShttle-hTRX转染Hela细胞系进行瞬时表达,通过Westem blot、免疫组化、活性测定、酶切、PCR扩增等方法证实构建载体的正确性.结果构建携带人硫氧还蛋白基因的真核表达载体pShttle-hTRX,转染HeLa细胞后可检测到hTRX的转录和表达.结论正确构建了携带人硫氧还蛋白基因的真核表达载体,本结果可用于探讨硫氧还蛋白的生物学作用。
Key concepts: Expression (computer science), Vector (molecular biology), Biology, Computational biology, Cell biology, Computer science, Genetics, Recombinant DNA