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目的 构建大鼠IL-10真核表达载体并在大鼠软骨细胞中进行表达。方法 将目的基因片段通过酶切连入真核表达载体pcDNA3,并以Super Fect Transfection Reagent介导转入大鼠软骨细胞表达,RT-PCR检测细胞中mRNA水平,及ELISA检测细胞培养上清液中的IL-10蛋白含量。结果 成功构建了大鼠IL-10真核表达载体,转入软骨细胞后,检测到细胞内IL-10mRNA转录,细胞培养24、48和72 h后,经ELISA检测上清液中IL-10蛋白含量分别为36、62和100 pg/mL。结论 大鼠IL-10真核表达载体的构建并在软骨细胞中表达,为研究IL-10诱导软骨细胞同种异体移植耐受奠定了基础。
Objective To construct a rat IL-10 eukaryotic expression vector and express it in rat chondrocytes. Methods The target gene fragment was inserted into the eukaryotic expression vector pcDNA3 by enzyme digestion, and then transfected into rat chondrocytes by Super Fect Transfection Reagent. The mRNA level in the cells was detected by RT-PCR, IL-10 protein content. Results The eukaryotic expression vector of rat IL-10 was successfully constructed. After transfected into chondrocytes, the intracellular IL-10 mRNA transcription was detected. After 24, 48 and 72 hours of cell culture, IL-10 protein The contents were 36, 62 and 100 pg / mL, respectively. Conclusion The construction of eukaryotic expression vector of rat IL-10 and its expression in chondrocytes lay the foundation for studying the tolerance of IL-10 to chondrocytes allograft.