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目的:通过转基因技术建立稳定表达hrCEMP1的成纤维细胞株。方法:利用高效率的阳离子聚合物,将含有hrCEMP1编码序列的真核表达质粒pcDNA3.1-hrCEMP1转染入NIH3T3细胞中,用G418筛选获得稳定转染细胞株,以RT-PCR检测hrCEMP1基因转录,以酶联免疫吸附试验(ELISA)检测hrCEMP1蛋白表达。结果:转染细胞株内有hrCEMP mRNA的转录,并可检测到hrCEMP1蛋白的表达。结论:通过转基因手段,可以将外源性hrCEMP1基因成功导入成纤维细胞内,为进一步研究hrCEMP1对成纤维细胞的诱导作用建立基础。
OBJECTIVE: To establish a fibroblast cell line stably expressing hrCEMP1 by transgenic technology. Methods: The eukaryotic expression plasmid pcDNA3.1-hrCEMP1 containing hrCEMP1 coding sequence was transfected into NIH3T3 cells by high efficiency cationic polymer. Stable transfected cell lines were screened by G418, and hrCEMP1 gene transcription was detected by RT-PCR The hrCEMP1 protein expression was detected by enzyme-linked immunosorbent assay (ELISA). Results: hrCEMP mRNA transcripts were detected in transfected cells and hrCEMP1 protein expression was detected. CONCLUSION: The exogenous hrCEMP1 gene can be successfully introduced into fibroblasts by transgenic means, so as to further study the induction of hrCEMP1 on fibroblasts.