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目的:为应用RNAi技术调节白细胞介素6(interleukin-6,IL-6)基因表达,构建大鼠IL-6基因真核表达质粒。方法:从大鼠结肠中提取总RNA,经逆转录(reverse transcription)得到目的基因的cDNA,经PCR得到目的基因扩增片段,将其连接入pcDNA3.1/CT-GFP-TOPO载体中,对重组体进行鉴定。结果:成功构建了目的基因的重组体,测序报告显示核苷酸序列无误。结论:大鼠真核表达质粒pcDNA3.1/CT-GFP-TOPO/IL-6的成功构建,为应用RNAi技术调节IL-6基因表达提供了实验基础,为研究IL-6与溃疡性结肠炎的关系提供实验平台。
OBJECTIVE: To construct an eukaryotic expression plasmid for rat IL-6 gene by RNAi technology to regulate the gene expression of interleukin-6 (IL-6). Methods: The total RNA was extracted from the colon of rats and the cDNA of the target gene was obtained by reverse transcription. The amplified fragment of the target gene was amplified by PCR and ligated into the pcDNA3.1 / CT-GFP-TOPO vector Recombinant identification. Results: The recombinants of the target gene were successfully constructed and the sequencing report showed that the nucleotide sequence was correct. CONCLUSION: The successful construction of rat eukaryotic expression plasmid pcDNA3.1 / CT-GFP-TOPO / IL-6 provided the experimental basis for the regulation of IL-6 gene expression using RNAi technology. In order to study the relationship between IL-6 and ulcerative colitis The relationship between the experimental platform.